Updated on 2022/05/11

写真a

 
SAKAGUCHI Masakiyo
 
Organization
Faculty of Medicine, Dentistry and Pharmaceutical Sciences Professor
Position
Professor
External link

Degree

  • 博士(医学) ( 岡山大学 )

Research Interests

  • 細胞生物学

  • Cell Biology

  • REIC/Dkk-3

  • S100タンパク質

  • がん

Research Areas

  • Life Science / Cell biology

Education

  • Okayama University   医学研究科   細胞生物学

    - 2001

      More details

    Country: Japan

    researchmap

  • Okayama University    

    - 2001

      More details

Research History

  • 岡山大学 学術研究院 医歯薬学域   教授

    2021.4

      More details

  • 岡山大学大学院医歯薬学総合研究科   教授

    2018.7 - 2020.3

      More details

  • - Professor,Graduate School of Medicine, Dentistry and Pharmaceutical Sciences,Okayama University

    2018

      More details

  • - 岡山大学医歯薬学総合研究科 教授

    2018

      More details

  • Associate Professor,Graduate School of Medicine, Dentistry and Pharmaceutical Sciences,Okayama University

    2008 - 2018

      More details

  • 岡山大学医歯薬学総合研究科 准教授

    2008 - 2018

      More details

  • Assistant Professor,Graduate School of Medicine, Dentistry and Pharmaceutical Sciences,Okayama University

    2007 - 2008

      More details

  • 岡山大学医歯薬学総合研究科 助教

    2007 - 2008

      More details

  • 岡山大学医歯薬学総合研究科 助手

    2005 - 2007

      More details

  • Research Associate,Graduate School of Medicine, Dentistry and Pharmaceutical Sciences,Okayama University

    2005 - 2007

      More details

  • 岡山大学医歯薬学総合研究科 助手

    2004 - 2005

      More details

  • Research Associate,Graduate School of Medicine, Dentistry and Pharmaceutical Sciences,Okayama University

    2004 - 2005

      More details

▼display all

Professional Memberships

▼display all

Committee Memberships

  • 日本組織培養学会   理事  

    2021.4   

      More details

  • 日本癌学会   評議員  

    2020.1   

      More details

  • 日本組織培養学会第90回大会   プログラム 運営委員  

    2017.7   

      More details

    Committee type:Academic society

    researchmap

  • 日本組織培養学会   大会運営補助  

    2004   

      More details

    Committee type:Academic society

    日本組織培養学会

    researchmap

 

Papers

  • Dkk3/REIC Deficiency Impairs Spermiation, Sperm Fibrous Sheath Integrity and the Sperm Motility of Mice. International journal

    Ruizhi Xue, Wenfeng Lin, Hirofumi Fujita, Jingkai Sun, Rie Kinoshita, Kazuhiko Ochiai, Junichiro Futami, Masami Watanabe, Hideyo Ohuchi, Masakiyo Sakaguchi, Zhengyan Tang, Peng Huang, Yasutomo Nasu, Hiromi Kumon

    Genes   13 ( 2 )   2022.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The role of Dickkopf-3 (Dkk3)/REIC (The Reduced Expression in Immortalized Cells), a Wnt-signaling inhibitor, in male reproductive physiology remains unknown thus far. To explore the functional details of Dkk3/REIC in the male reproductive process, we studied the Dkk3/REIC knock-out (KO) mouse model. By examining testicular sections and investigating the sperm characteristics (count, vitality and motility) and ultrastructure, we compared the reproductive features between Dkk3/REIC-KO and wild-type (WT) male mice. To further explore the underlying molecular mechanism, we performed RNA sequencing (RNA-seq) analysis of testicular tissues. Our results showed that spermiation failure existed in seminiferous tubules of Dkk3/REIC-KO mice, and sperm from Dkk3/REIC-KO mice exhibited inferior motility (44.09 ± 8.12% vs. 23.26 ± 10.02%, p < 0.01). The Ultrastructure examination revealed defects in the sperm fibrous sheath of KO mice. Although the average count of Dkk3/REIC-KO epididymal sperm was less than that of the wild-types (9.30 ± 0.69 vs. 8.27 ± 0.87, ×106), neither the gap (p > 0.05) nor the difference in the sperm vitality rate (72.83 ± 1.55% vs. 72.50 ± 0.71%, p > 0.05) were statistically significant. The RNA-seq and GO (Gene Oncology) enrichment results indicated that the differential genes were significantly enriched in the GO terms of cytoskeleton function, cAMP signaling and calcium ion binding. Collectively, our research demonstrates that Dkk3/REIC is involved in the process of spermiation, fibrous sheath integrity maintenance and sperm motility of mice.

    DOI: 10.3390/genes13020285

    PubMed

    researchmap

  • Multifaceted Analysis of IL-23A- and/or EBI3-Including Cytokines Produced by Psoriatic Keratinocytes. International journal

    Kota Tachibana, Nina Tang, Hitoshi Urakami, Ai Kajita, Mina Kobashi, Hayato Nomura, Minori Sasakura, Satoru Sugihara, Fan Jiang, Nahoko Tomonobu, Masakiyo Sakaguchi, Mamoru Ouchida, Shin Morizane

    International journal of molecular sciences   22 ( 23 )   2021.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Interleukin (IL) 23 (p19/p40) plays a critical role in the pathogenesis of psoriasis and is upregulated in psoriasis skin lesions. In clinical practice, anti-IL-23Ap19 antibodies are highly effective against psoriasis. IL-39 (p19/ Epstein-Barr virus-induced (EBI) 3), a newly discovered cytokine in 2015, shares the p19 subunit with IL-23. Anti-IL-23Ap19 antibodies may bind to IL-39; also, the cytokine may contribute to the pathogenesis of psoriasis. To investigate IL23Ap19- and/or EBI3-including cytokines in psoriatic keratinocytes, we analyzed IL-23Ap19 and EBI3 expressions in psoriasis skin lesions, using immunohistochemistry and normal human epidermal keratinocytes (NHEKs) stimulated with inflammatory cytokines, using quantitative real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and liquid chromatography-electrospray tandem mass spectrometry (LC-Ms/Ms). Immunohistochemical analysis showed that IL-23Ap19 and EBI3 expressions were upregulated in the psoriasis skin lesions. In vitro, these expressions were synergistically induced by the triple combination of tumor necrosis factor (TNF)-α, IL-17A, and interferon (IFN)-γ, and suppressed by dexamethasone, vitamin D3, and acitretin. In ELISA and LC-Ms/Ms analyses, keratinocyte-derived IL-23Ap19 and EBI3, but not heterodimeric forms, were detected with humanized anti-IL-23Ap19 monoclonal antibodies, tildrakizumab, and anti-EBI3 antibodies, respectively. Psoriatic keratinocytes may express IL-23Ap19 and EBI3 proteins in a monomer or homopolymer, such as homodimer or homotrimer.

    DOI: 10.3390/ijms222312659

    PubMed

    researchmap

  • Oncogenic potential of human pluripotent stem cell-derived lung organoids with HER2 overexpression. Reviewed International journal

    Akihiro Miura, Daisuke Yamada, Masahiro Nakamura, Shuta Tomida, Dai Shimizu, Yan Jiang, Tomoka Takao, Hiromasa Yamamoto, Ken Suzawa, Kazuhiko Shien, Masaomi Yamane, Masakiyo Sakaguchi, Shinichi Toyooka, Takeshi Takarada

    International journal of cancer   149 ( 8 )   1593 - 1604   2021.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Lung adenocarcinoma (LUAD) is the most common types among lung cancers generally arising from terminal airway and understanding of multistep carcinogenesis is crucial to develop novel therapeutic strategy for LUAD. Here we used human induced pluripotent stem cells (hiPSCs) to establish iHER2-hiPSCs in which doxycycline induced the expression of the oncoprotein human epidermal growth factor receptor 2 (HER2)/ERBB2. Lung progenitors that differentiated from iHER2-hiPSCs, which expressed NKX2-1/TTF-1 known as a lung lineage maker, were cocultured with human fetal fibroblast and formed human lung organoids (HLOs) comprising alveolar type 2-like cells. HLOs that overexpressed HER2 transformed to tumor-like structures similar to atypical adenomatous hyperplasia, which is known for lung precancerous lesion and upregulated the activities of oncogenic signaling cascades such as RAS/RAF/MAPK and PI3K/AKT/mTOR. The degree of morphological irregularity and proliferation capacity were significantly higher in HLOs from iHER2-hiPSCs. Moreover, the transcriptome profile of the HLOs shifted from a normal lung tissue-like state to one characteristic of clinical LUAD with HER2 amplification. Our results suggest that hiPSC-derived HLOs may serve as a model to recapitulate the early tumorigenesis of LUAD and would provide new insights into the molecular basis of tumor initiation and progression.

    DOI: 10.1002/ijc.33713

    PubMed

    researchmap

  • Inhibitory effects of RAGE-aptamer on development of monocrotaline-induced pulmonary arterial hypertension in rats. Reviewed International journal

    Kazufumi Nakamura, Satoshi Akagi, Kentaro Ejiri, Masashi Yoshida, Toru Miyoshi, Masakiyo Sakaguchi, Naofumi Amioka, Luh Oliva Saraswati Suastika, Megumi Kondo, Rie Nakayama, Yoichi Takaya, Yuichiro Higashimoto, Kei Fukami, Hiromi Matsubara, Hiroshi Ito

    Journal of cardiology   78 ( 1 )   12 - 16   2021.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    BACKGROUND: The receptor for advanced glycation end products (RAGE), a transmembrane receptor belonging to the immunoglobulin superfamily, is overexpressed in pulmonary artery smooth muscle cells (PASMCs) in patients with pulmonary arterial hypertension (PAH) and is implicated in the etiology of PAH. Recently, we reported that RAGE-aptamer, a short and single-stranded DNA directed against RAGE, inhibited an inappropriate increase in cultured PASMCs in PAH. The aim of this study was to determine the efficacy of RAGE-aptamer in monocrotaline-induced PAH in rats. METHODS AND RESULTS: Rats were assigned to either an untreated control group, a group that received continuous subcutaneous administration of RAGE-aptamer immediately after monocrotaline injection, or a group that received control-aptamer immediately after monocrotaline injection. All rats survived 21 days after injection of monocrotaline and control-aptamer or RAGE-aptamer. Injection of monocrotaline with continuous subcutaneous delivery of control-aptamer resulted in higher right ventricular systolic pressure compared with controls. This increase was attenuated by continuous subcutaneous delivery of RAGE-aptamer. The proportion of small pulmonary arteries with full muscularization was greater in the monocrotaline and control-aptamer group than in the control group. Continuous subcutaneous delivery of RAGE-aptamer significantly reduced the percentage of small pulmonary arteries with full muscularization. CONCLUSIONS: Continuous subcutaneous delivery of RAGE-aptamer suppresses development of monocrotaline-induced PAH in rats. Inhibition of RAGE ameliorates muscularization of small pulmonary arteries. Treatment with RAGE-aptamer might be a new therapeutic option for PAH.

    DOI: 10.1016/j.jjcc.2020.12.009

    PubMed

    researchmap

  • PLOD2-driven IL-6/STAT3 signaling promotes the invasion and metastasis of oral squamous cell carcinoma via activation of integrin β1. Reviewed International journal

    Ken Saito, Ayaka Mitsui, I Wayan Sumardika, Yusuke Yokoyama, Masakiyo Sakaguchi, Eisaku Kondo

    International journal of oncology   58 ( 6 )   2021.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We previously reported that high expression of procollagen‑lysine 2‑oxoglutarate 5‑dioxygenase 2 (PLOD2) leads to stabilization and plasma membrane translocation of integrin β1 to promote the invasion and metastasis of oral squamous cell carcinoma (SCC). The present study aimed to further understand the relationship between PLOD2‑integrin β1 signaling and the tumor microenvironment. This study provided further advanced insights indicating that elevated interleukin (IL)‑6 in the tumor microenvironment acts as a key molecule that triggers PLOD2‑integrin β1 axis‑derived acceleration of tumor invasion and metastasis. It was found using the dual‑luciferase reporter assay system that signal transducer and activator of transcription 3 (STAT3) activation by IL‑6 was essential for increasing the expression levels of PLOD2 through direct activation of the PLOD2 promoter in oral SCC, whereas IL‑6 stimulation did not contribute to integrin β1 expression or the subsequent maturation process towards a functional form on the plasma membrane. Furthermore, the expression of IL‑6 in oral SCC tissues was mainly observed in the tumor stroma. Finally, with double immunofluorescence staining, it was found that IL‑6 expression occurred in CD163‑positive M2 macrophages distributed around the tumor nest. These results combined with our previous results indicate that as IL‑6 significantly increases STAT3‑mediated PLOD2 promoter activity, IL‑6 released by M2‑type tumor‑associated macrophages is a crucial factor that promotes PLOD2‑integrin β1 axis‑enhanced invasion and metastasis of oral SCC cells.

    DOI: 10.3892/ijo.2021.5209

    PubMed

    researchmap

  • Presence of Microplastics in Four Types of Shellfish Purchased at Fish Markets in Okayama City, Japan.

    Kenichi Yamamoto, Toshiyuki Oshiki, Hiroko Kagawa, Masayoshi Namba, Masakiyo Sakaguchi

    Acta medica Okayama   75 ( 3 )   381 - 384   2021.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The worldwide microplastic pollution in our environment is a matter of great concern. Harmful effects of plastics have been reported in various types of organisms including murine animals. We examined the presence of microplastics in four types of shellfish purchased from fish markets in Okayama, Japan and served to the public: short-neck clam (Ruditapes philippinarum, asari in Japanese), hard-shell clam (Meretrix lusoria, hamaguri), brackishwater clam (Cyrenidae, shijimi), and oyster (Crassostrea gigas, kaki). Our analyses demonstrated that approx. 3 pieces of microplastics were present per single shellfish, based on the division of the total number of pieces of microplastic obtained from all 4 types of shellfish by the total number of shellfish examined. Since health problems in humans due to microplastics have not yet been confirmed, further examinations of the effects of ingested microplastics are needed.

    DOI: 10.18926/AMO/62234

    PubMed

    researchmap

  • Histidine-Rich Glycoprotein Stimulates Human Neutrophil Phagocytosis and Prolongs Survival through CLEC1A. Reviewed International journal

    Yohei Takahashi, Hidenori Wake, Masakiyo Sakaguchi, Yukinori Yoshii, Kiyoshi Teshigawara, Dengli Wang, Masahiro Nishibori

    Journal of immunology (Baltimore, Md. : 1950)   206 ( 4 )   737 - 750   2021.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Histidine-rich glycoprotein (HRG) is a multifunctional plasma protein and maintains the homeostasis of blood cells and vascular endothelial cells. In the current study, we demonstrate that HRG and recombinant HRG concentration dependently induced the phagocytic activity of isolated human neutrophils against fluorescence-labeled Escherichia coli and Staphylococcus aureus through the stimulation of CLEC1A receptors, maintaining their spherical round shape. The phagocytosis-inducing effects of HRG were inhibited by a specific anti-HRG Ab and enhanced by opsonization of bacteria with diluted serum. HRG and C5a prolonged the survival time of isolated human neutrophils, in association with a reduction in the spontaneous production of extracellular ROS. In contrast, HRG maintained the responsiveness of neutrophils to TNF-α, zymosan, and E. coli with regard to reactive oxygen species production. The blocking Ab for CLEC1A and recombinant CLEC1A-Fc fusion protein significantly inhibited the HRG-induced neutrophil rounding, phagocytic activity, and prolongation of survival time, suggesting the involvement of the CLEC1A receptor in the action of HRG on human neutrophils. These results as a whole indicated that HRG facilitated the clearance of E. coli and S. aureus by maintaining the neutrophil morphology and phagocytosis, contributing to the antiseptic effects of HRG in vivo.

    DOI: 10.4049/jimmunol.2000817

    PubMed

    researchmap

  • The heterodimer S100A8/A9 is a potent therapeutic target for idiopathic pulmonary fibrosis. Reviewed International journal

    Kota Araki, Rie Kinoshita, Nahoko Tomonobu, Yuma Gohara, Shuta Tomida, Yuta Takahashi, Satoru Senoo, Akihiko Taniguchi, Junko Itano, Ken-Ichi Yamamoto, Hitoshi Murata, Ken Suzawa, Kazuhiko Shien, Hiromasa Yamamoto, Mikio Okazaki, Seiichiro Sugimoto, Kouichi Ichimura, Masahiro Nishibori, Nobuaki Miyahara, Shinichi Toyooka, Masakiyo Sakaguchi

    Journal of molecular medicine (Berlin, Germany)   99 ( 1 )   131 - 145   2021.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    In patients with interstitial pneumonia, pulmonary fibrosis is an irreversible condition that can cause respiratory failure. Novel treatments for pulmonary fibrosis are necessary. Inflammation is thought to activate lung fibroblasts, resulting in pulmonary fibrosis. Of the known inflammatory molecules, we have focused on S100A8/A9 from the onset of inflammation to the subsequent progression of inflammation. Our findings confirmed the high expression of S100A8/A9 in specimens from patients with pulmonary fibrosis. An active role of S100A8/A9 was demonstrated not only in the proliferation of fibroblasts but also in the fibroblasts' differentiation to myofibroblasts (the active form of fibroblasts). S100A8/A9 also forced fibroblasts to upregulate the production of collagen. These effects were induced via the receptor of S100A8/A9, i.e., the receptor for advanced glycation end products (RAGE), on fibroblasts. The anti-S100A8/A9 neutralizing antibody inhibited the effects of S100A8/A9 on fibroblasts and suppressed the progression of fibrosis in bleomycin (BLM)-induced pulmonary fibrosis mouse model. Our findings strongly suggest a crucial role of S100A8/A9 in pulmonary fibrosis and the usefulness of S100A8/A9-targeting therapy for fibrosis interstitial pneumonia. HIGHLIGHTS: S100A8/A9 level is highly upregulated in the IPF patients' lungs as well as the blood. S100A8/A9 promotes not only the growth of fibroblasts but also differentiation to myofibroblasts. The cell surface RAGE acts as a crucial receptor to the extracellular S100A8/A9 in fibroblasts. The anti-S100A8/A9 antibody effectively suppresses the progression of IPF in a mouse model. In idiopathic pulmonary fibrosis (IPF), S100A8/A9, a heterodimer composed of S100A8 and S100A9 proteins, plays a crucial role in the onset of inflammation and the subsequent formation of a feed-forward inflammatory loop that promotes fibrosis. (1) The local, pronounced increase in S100A8/A9 in the injured inflammatory lung region-which is provided mainly by the activated neutrophils and macrophages-exerts strong inflammatory signals accompanied by dozens of inflammatory soluble factors including cytokines, chemokines, and growth factors that further act to produce and secrete S100A8/A9, eventually making a sustainable inflammatory circuit that supplies an indefinite presence of S100A8/A9 in the extracellular space with a mal-increased level. (2) The elevated S100A8/A9 compels fibroblasts to activate through receptor for advanced glycation end products (RAGE), one of the major S100A8/A9 receptors, resulting in the activation of NFκB, leading to fibroblast mal-events (e.g., elevated cell proliferation and transdifferentiation to myofibroblasts) that actively produce not only inflammatory cytokines but also collagen matrices. (3) Finally, the S100A8/A9-derived activation of lung fibroblasts under a chronic inflammation state leads to fibrosis events and constantly worsens fibrosis in the lung. Taken together, these findings suggest that the extracellular S100A8/A9 heterodimer protein is a novel mainstay soluble factor for IPF that exerts many functions as described above (1-3). Against this background, we herein applied the developed S100A8/A9 neutralizing antibody to prevent IPF. The IPF imitating lung fibrosis in an IPF mouse model was effectively blocked by treatment with the antibody, leading to enhanced survival. The developed S100A8/A9 antibody, as an innovative novel biologic, may help shed light on the difficulties encountered with IPF therapy in clinical settings.

    DOI: 10.1007/s00109-020-02001-x

    PubMed

    researchmap

  • RUNX2 Phosphorylation by Tyrosine Kinase ABL Promotes Breast Cancer Invasion. Reviewed International journal

    Fang He, Yoshinori Matsumoto, Yosuke Asano, Yuriko Yamamura, Takayuki Katsuyama, Jose La Rose, Nahoko Tomonobu, Ni Luh Gede Yoni Komalasari, Masakiyo Sakaguchi, Robert Rottapel, Jun Wada

    Frontiers in oncology   11   665273 - 665273   2021

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Activity of transcription factors is normally regulated through interaction with other transcription factors, chromatin remodeling proteins and transcriptional co-activators. In distinction to these well-established transcriptional controls of gene expression, we have uncovered a unique activation model of transcription factors between tyrosine kinase ABL and RUNX2, an osteoblastic master transcription factor, for cancer invasion. We show that ABL directly binds to, phosphorylates, and activates RUNX2 through its SH2 domain in a kinase activity-dependent manner and that the complex formation of these proteins is required for expression of its target gene MMP13. Additionally, we show that the RUNX2 transcriptional activity is dependent on the number of its tyrosine residues that are phosphorylated by ABL. In addition to regulation of RUNX2 activity, we show that ABL transcriptionally enhances RUNX2 expression through activation of the bone morphogenetic protein (BMP)-SMAD pathway. Lastly, we show that ABL expression in highly metastatic breast cancer MDA-MB231 cells is associated with their invasive capacity and that ABL-mediated invasion is abolished by depletion of endogenous RUNX2 or MMP13. Our genetic and biochemical evidence obtained in this study contributes to a mechanistic insight linking ABL-mediated phosphorylation and activation of RUNX2 to induction of MMP13, which underlies a fundamental invasive capacity in cancer and is different from the previously described model of transcriptional activation.

    DOI: 10.3389/fonc.2021.665273

    PubMed

    researchmap

  • Corrigendum: RUNX2 Phosphorylation by Tyrosine Kinase ABL Promotes Breast Cancer Invasion. International journal

    Fang He, Yoshinori Matsumoto, Yosuke Asano, Yuriko Yamamura, Takayuki Katsuyama, Jose La Rose, Nahoko Tomonobu, Ni Luh Gede Yoni Komalasari, Masakiyo Sakaguchi, Robert Rottapel, Jun Wada

    Frontiers in oncology   11   729192 - 729192   2021

     More details

    Language:English  

    [This corrects the article DOI: 10.3389/fonc.2021.665273.].

    DOI: 10.3389/fonc.2021.729192

    PubMed

    researchmap

  • Analysis of a plasmid encoding botulinum neurotoxin type G gene in Clostridium argentinense. Reviewed International journal

    Yoshihiko Sakaguchi, Jumpei Uchiyama, Akira Také, Kazuyoshi Gotoh, Masakiyo Sakaguchi, Tomonori Suzuki, Yumiko Yamamoto, Koji Hosomi, Tomoko Kohda, Masafumi Mukamoto, Shunji Kozaki, Shunji Hayashi, Keiji Oguma

    Anaerobe   66   102281 - 102281   2020.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Clostridium argentinense produces botulinum neurotoxin type G (BoNT/G). We sequenced and analyzed the plasmid harboring the bont/G gene, designated pCAG, in C. argentinense strain 2740. The pCAG consisted of 140,070 bp containing the bont/G gene cluster. Although this gene cluster showed high similarities in its DNA sequence and ORF arrangement to those of other bont gene clusters, the other regions of the plasmid did not. A phylogenetic study suggested that pCAG had a unique evolutionary history compared with other clostridial bont-harboring plasmids. This suggests that pCAG is possibly a novel type of plasmid expressing the bont/G gene in C. argentinense.

    DOI: 10.1016/j.anaerobe.2020.102281

    PubMed

    researchmap

  • Enhancement of intestinal epithelial barrier function by Weissella confusa F213 and Lactobacillus rhamnosus FBB81 probiotic candidates in an in vitro model of hydrogen peroxide-induced inflammatory bowel disease. Reviewed International journal

    Ni Nengah Dwi Fatmawati, Kazuyoshi Gotoh, I Putu Bayu Mayura, Komang Ayu Nocianitri, Gede Ngurah Rsi Suwardana, Ni Luh Gede Yoni Komalasari, Yan Ramona, Masakiyo Sakaguchi, Osamu Matsushita, I Nengah Sujaya

    BMC research notes   13 ( 1 )   489 - 489   2020.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    OBJECTIVE: Weissella confusa F213 (WCF213) and Lactobacillus rhamnosus FBB81 (LrFBB81) are two probiotic candidates isolated from humans in our previous study. Their functional activity on the mucosal barrier has not yet been adequately investigated. Therefore, the objective of this study was to investigate the effect of these strains on maintaining mucosal integrity in vitro. Caco-2 cell monolayers were pretreated with WCF213 and LrFBB81 before being exposed to hydrogen peroxide. The integrity of mucosal cells was evaluated by measuring the transepithelial resistance (TER), flux of FITC-labelled dextran, and ZO-1 protein distribution with the help of an immunofluorescence method. RESULTS: WCF213 was found to significantly maintain the TER better than the control hydrogen peroxide-treated cells (p < 0.001), followed by the strain combination, and LrFBB81 alone (p < 0.05). The permeability of mucosa was also successfully maintained by the WCF213 strain. This was illustrated by the significant reduction in the flux of FITC-labelled dextran (p < 0.05), which was larger than that exhibited by the other groups. The ZO-1 distribution of strain-treated cells showed less disruption than hydrogen peroxide-treated cells, consistent with the TER and FITC experimental results. These findings indicate that WCF213 and LrFBB81 plays important roles in the maintenance of mucosal integrity in a strain-dependent manner.

    DOI: 10.1186/s13104-020-05338-1

    PubMed

    researchmap

  • Synergistic regulation of hepatic Fsp27b expression by HNF4α and CREBH. Reviewed International journal

    Carlos Ichiro Kasano-Camones, Masayuki Takizawa, Wakana Iwasaki, Shota Sasaki, Mume Hamada, Aoi Morimoto, Masakiyo Sakaguchi, Frank J Gonzalez, Yusuke Inoue

    Biochemical and biophysical research communications   530 ( 2 )   432 - 439   2020.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The CIDE (cell death-inducing DFF45-like effector) family composed of CIDEA, CIDEB, CIDEC/FSP27 (fat-specific protein 27), has a critical role in growth of lipid droplets. Of these, CIDEB and CIDEC2/FSP27B are abundant in the liver, and the steatotic livers, respectively. Hepatocyte nuclear factor 4α (HNF4α) has an important role in lipid homeostasis because liver-specific HNF4α-null mice (Hnf4aΔHep mice) exhibit hepatosteatosis. We investigated whether HNF4α directly regulates expression of CIDE family genes. Expression of Cideb and Fsp27b was largely decreased in Hnf4aΔHep mice, while expression of Cidea was increased. Similar results were observed only in CIDEC2, the human orthologue of the Fsp27b, in human hepatoma cell lines in which HNF4α expression was knocked down. Conversely, overexpression of HNF4α strongly induced CIDEC2 expression in hepatoma cell lines. Furthermore, HNF4α transactivated Fsp27b by direct binding to an HNF4α response element in the Fsp27b promoter. In addition, Fsp27b is known to be transactivated by CREBH that is regulated by HNF4α, and expression of CREBH was induced by HNF4α in human hepatoma cells. Co-transfection of HNF4α and CREBH resulted in synergistic transactivation and induction of Fsp27b compared to that of HNF4α or CREBH alone. These results suggest that HNF4α, in conjunction with CREBH, plays an important role in regulation of Fsp27b expression.

    DOI: 10.1016/j.bbrc.2020.05.070

    PubMed

    researchmap

  • Cytotoxic Effects of Alcohol Extracts from a Plastic Wrap (Polyvinylidene Chloride) on Human Cultured Liver Cells and Mouse Primary Cultured Liver Cells. Reviewed

    Ken-Ichi Yamamoto, Hiroko Kagawa, Sakae Arimoto, Xian Wen Tan, Kento Yasui, Toshiyuki Oshiki, Masakiyo Sakaguchi

    Acta medica Okayama   74 ( 4 )   327 - 334   2020.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    An increasing accumulation of microplastics and further degraded nanoplastics in our environment is suspected to have harmful effects on humans and animals. To clarify this problem, we tested the cytotoxicity of two types of plastic wrap on human cultured liver cells and mouse primary cultured liver cells. Alcohol extracts from plastic wrap, i.e., polyvinylidene chloride (PVDC), showed cytotoxic effects on the cells. Alcohol extracts of polyethylene (PE) wrap were not toxic. The commercially available PVDC wrap consists of vinylidene chloride, epoxidized soybean oil, epoxidized linseed oil as a stiffener and stabilizer; we sought to identify which component(s) are toxic. The epoxidized soybean oil and epoxidized linseed oil exerted strong cytotoxicity, but the plastic raw material itself, vinylidene chloride, did not. Our findings indicate that plastic wraps should be used with caution in order to prevent health risks.

    DOI: 10.18926/AMO/60371

    PubMed

    researchmap

  • Neuroplastinβ-mediated upregulation of solute carrier family 22 member 18 antisense (SLC22A18AS) plays a crucial role in the epithelial-mesenchymal transition, leading to lung cancer cells' enhanced motility. Reviewed International journal

    Karolina Bajkowska, I Wayan Sumardika, Nahoko Tomonobu, Youyi Chen, Ken-Ichi Yamamoto, Rie Kinoshita, Hitoshi Murata, Ni Luh Gede Yoni Komalasari, Fan Jiang, Akira Yamauchi, I Made Winarsa Ruma, Carlos Ichiro Kasano-Camones, Yusuke Inoue, Masakiyo Sakaguchi

    Biochemistry and biophysics reports   22   100768 - 100768   2020.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    Our recent study revealed an important role of the neuroplastin (NPTN)β downstream signal in lung cancer dissemination in the lung. The molecular mechanism of the signal pathway downstream of NPTNβ is a serial activation of the key molecules we identified: tumor necrosis factor (TNF) receptor-associated factor 2 (TRAF2) adaptor, nuclear factor (NF)IA/NFIB heterodimer transcription factor, and SAM pointed-domain containing ETS transcription factor (SPDEF). The question of how dissemination is controlled by SPDEF under the activated NPTNβ has not been answered. Here, we show that the NPTNβ-SPDEF-mediated induction of solute carrier family 22 member 18 antisense (SLC22A18AS) is definitely required for the epithelial-mesenchymal transition (EMT) through the NPTNβ pathway in lung cancer cells. In vitro, the induced EMT is linked to the acquisition of active cellular motility but not growth, and this is correlated with highly disseminative tumor progression in vivo. The publicly available data also show the poor survival of SLC22A18AS-overexpressing lung cancer patients. Taken together, these data highlight a crucial role of SLC22A18AS in lung cancer dissemination, which provides novel input of this molecule to the signal cascade of NPTNβ. Our findings contribute to a better understanding of NPTNβ-mediated lung cancer metastasis.

    DOI: 10.1016/j.bbrep.2020.100768

    PubMed

    researchmap

  • Histidine-Rich Glycoprotein Inhibits High-Mobility Group Box-1-Mediated Pathways in Vascular Endothelial Cells through CLEC-1A. Reviewed International journal

    Shangze Gao, Hidenori Wake, Masakiyo Sakaguchi, Dengli Wang, Youhei Takahashi, Kiyoshi Teshigawara, Hui Zhong, Shuji Mori, Keyue Liu, Hideo Takahashi, Masahiro Nishibori

    iScience   23 ( 6 )   101180 - 101180   2020.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    High-mobility group box-1 (HMGB1) protein has been postulated to play a pathogenic role in severe sepsis. Histidine-rich glycoprotein (HRG), a 75 kDa plasma protein, was demonstrated to improve the survival rate of septic mice through the regulation of neutrophils and endothelium barrier function. As the relationship of HRG and HMGB1 remains poorly understood, we investigated the effects of HRG on HMGB1-mediated pathway in endothelial cells, focusing on the involvement of specific receptors for HRG. HRG potently inhibited the HMGB1 mobilization and effectively suppressed rHMGB1-induced inflammatory responses and expression of all three HMGB1 receptors in endothelial cells. Moreover, we first clarified that these protective effects of HRG on endothelial cells were mediated through C-type lectin domain family 1 member A (CLEC-1A) receptor. Thus, current study elucidates protective effects of HRG on vascular endothelial cells through inhibition of HMGB1-mediated pathways may contribute to the therapeutic effects of HRG on severe sepsis.

    DOI: 10.1016/j.isci.2020.101180

    PubMed

    researchmap

  • MAPK Pathway Alterations Correlate with Poor Survival and Drive Resistance to Therapy in Patients with Lung Cancers Driven by ROS1 Fusions. Reviewed International journal

    Hiroki Sato, Adam J Schoenfeld, Evan Siau, Yue Christine Lu, Huichun Tai, Ken Suzawa, Daisuke Kubota, Allan J W Lui, Besnik Qeriqi, Marissa Mattar, Michael Offin, Masakiyo Sakaguchi, Shinichi Toyooka, Alexander Drilon, Neal X Rosen, Mark G Kris, David Solit, Elisa De Stanchina, Monika A Davare, Gregory J Riely, Marc Ladanyi, Romel Somwar

    Clinical cancer research : an official journal of the American Association for Cancer Research   26 ( 12 )   2932 - 2945   2020.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    PURPOSE: ROS1 tyrosine kinase inhibitors (TKI) provide significant benefit in lung adenocarcinoma patients with ROS1 fusions. However, as observed with all targeted therapies, resistance arises. Detecting mechanisms of acquired resistance (AR) is crucial to finding novel therapies and improve patient outcomes. EXPERIMENTAL DESIGN: ROS1 fusions were expressed in HBEC and NIH-3T3 cells either by cDNA overexpression (CD74/ROS1, SLC34A2/ROS1) or CRISPR-Cas9-mediated genomic engineering (EZR/ROS1). We reviewed targeted large-panel sequencing data (using the MSK-IMPACT assay) patients treated with ROS1 TKIs, and genetic alterations hypothesized to confer AR were modeled in these cell lines. RESULTS: Eight of the 75 patients with a ROS1 fusion had a concurrent MAPK pathway alteration and this correlated with shorter overall survival. In addition, the induction of ROS1 fusions stimulated activation of MEK/ERK signaling with minimal effects on AKT signaling, suggesting the importance of the MAPK pathway in driving ROS1 fusion-positive cancers. Of 8 patients, 2 patients harbored novel in-frame deletions in MEK1 (MEK1delE41_L54) and MEKK1 (MEKK1delH907_C916) that were acquired after ROS1 TKIs, and 2 patients harbored NF1 loss-of-function mutations. Expression of MEK1del or MEKK1del, and knockdown of NF1 in ROS1 fusion-positive cells activated MEK/ERK signaling and conferred resistance to ROS1 TKIs. Combined targeting of ROS1 and MEK inhibited growth of cells expressing both ROS1 fusion and MEK1del. CONCLUSIONS: We demonstrate that downstream activation of the MAPK pathway can mediate of innate acquired resistance to ROS1 TKIs and that patients harboring ROS1 fusion and concurrent downstream MAPK pathway alterations have worse survival. Our findings suggest a treatment strategy to target both aberrations.

    DOI: 10.1158/1078-0432.CCR-19-3321

    PubMed

    researchmap

  • Xylitol acts as an anticancer monosaccharide to induce selective cancer death via regulation of the glutathione level. Reviewed International journal

    Nahoko Tomonobu, Ni Luh Gede Yoni Komalasari, I Wayan Sumardika, Fan Jiang, Youyi Chen, Ken-Ichi Yamamoto, Rie Kinoshita, Hitoshi Murata, Yusuke Inoue, Masakiyo Sakaguchi

    Chemico-biological interactions   324   109085 - 109085   2020.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Herbal medicines and their bioactive compounds are increasingly being recognized as useful drugs for cancer treatments. The parasitic fungus Cordyceps militaris is an attractive anticancer herbal since it shows very powerful anticancer activity due to its phytocompound cordycepin. We previously discovered and reported that a high amount of xylitol is present in Cordyceps militaris extract, and that xylitol unexpectedly showed anticancer activity in a cancer-selective manner. We thus hypothesized that xylitol could become a useful supplement to help prevent various cancers, if we can clarify the specific machinery by which xylitol induces cancer cell death. It is also unclear whether xylitol acts on cancer suppression in vivo as well as in vitro. Here we show for the first time that induction of the glutathione-degrading enzyme CHAC1 is the main cause of xylitol-induced apoptotic cell death in cancer cells. The induction of CHAC1 is required for the endoplasmic reticulum (ER) stress that is triggered by xylitol in cancer cells, and is linked to a second induction of oxidative stress in the treated cells, and eventually leads to apoptotic cell death. Our in vivo approach also demonstrated that an intravenous injection of xylitol had a tumor-suppressing effect in mice, to which the xylitol-triggered ER stress also greatly contributed. We also observed that xylitol efficiently sensitized cancer cells to chemotherapeutic drugs. Based on our findings, a chemotherapeutic strategy combined with xylitol might improve the outcomes of patients facing cancer.

    DOI: 10.1016/j.cbi.2020.109085

    PubMed

    researchmap

  • exMCAM-Fc, an S100A8/A9-mediated-metastasis blocker, efficiently reduced the number of circulating tumor cells that appeared in the blood flow. Reviewed International journal

    Nahoko Tomonobu, Rie Kinoshita, Masakiyo Sakaguchi

    Molecular biology reports   47 ( 6 )   4879 - 4883   2020.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Metastasis is the major cause of treatment failure in cancer patients and of cancer-associated death so that therapeutic regulation of metastasis is very important subject for the cancer treatment. We have been reported that S100A8/A9, a heterodimer complex of S100A8 and S100A9, and its receptors play a crucial role in the lung tropic cancer metastasis, i.e., S100A8/A9 is actively secreted from the lung when cancer mass exists even at remote area from the lung and then functions to attract the distant cancer cells to the lung since cancer cells own the S100A8/A9 receptor(s) on their cell surface. Interestingly, one of the newly developed decoys, exMCAM-Fc, a Fc fusion protein with the extracellular region of melanoma cell adhesion molecule (MCAM), one of the S100A8/A9 receptors, that could prevent the interaction of S100A8/A9 with MCAM, efficiently suppressed the lung tropic cancer metastasis through exerting the several inhibitory effects on the S100A8/A9-mediated cancer cell events including enhanced mobility, invasion and attachment to the endothelial cells. However, it still remains to clarify if the decoy will reduce the number of circulating tumor cells (CTCs) that are defined as substantial cells in the context of organ tropic cancer metastasis. Here, we first show that exMCAM-Fc effectively reduces the number of CTCs in the blood flow of the melanoma bearing mice. The novel finding reinforces the suppressive role of exMCAM-Fc on the cancer metastasis. We therefore expect that exMCAM-Fc may greatly contribute to reduce treatment failure by the efficient blocking of the life threatening cancer metastasis.

    DOI: 10.1007/s11033-020-05495-3

    PubMed

    researchmap

  • Antitumor Effects of Pan-RAF Inhibitor LY3009120 Against Lung Cancer Cells Harboring Oncogenic BRAF Mutation. Reviewed International journal

    Shunsaku Miyauchi, Kazuhiko Shien, Tatsuaki Takeda, Kota Araki, Kentaro Nakata, Akihiro Miura, Yuta Takahashi, Eisuke Kurihara, Yusuke Ogoshi, Kei Namba, Ken Suzawa, Hiromasa Yamamoto, Mikio Okazaki, Junichi Soh, Shuta Tomida, Masaomi Yamane, Masakiyo Sakaguchi, Shinichi Toyooka

    Anticancer research   40 ( 5 )   2667 - 2673   2020.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    BACKGROUND/AIM: The therapeutic strategy for patients with non-small-cell lung cancer (NSCLC) harboring the BRAF non-V600E mutation has not been established. LY3009120, a newly discovered pan-RAF inhibitor, has shown strong antitumor effects in cancers with various BRAF genotypes. This study investigated the antitumor effects of LY3009120 in NSCLC cells harboring the BRAF non-V600E mutation. MATERIALS AND METHODS: We examined the antitumor effects of LY3009120 by MTS assay and flow cytometry. We analyzed the expression status of proteins by western blot. The mouse xenograft models were used for the in vivo experiments. RESULTS: LY3009120 suppressed BRAF-related downstream pathway molecules and induced cleavage of poly ADP-ribose polymerase in all examined NSCLC cell lines. LY3009120 also inhibited in vivo tumor growth in NSCLC cells harboring the BRAF non-V600E mutation. CONCLUSION: LY3009120 is a potent therapeutic agent for patients with BRAF non-V600E mutant NSCLC.

    DOI: 10.21873/anticanres.14237

    PubMed

    researchmap

  • S100 Soil Sensor Receptors and Molecular Targeting Therapy Against Them in Cancer Metastasis. Reviewed International journal

    Nahoko Tomonobu, Rie Kinoshita, Masakiyo Sakaguchi

    Translational oncology   13 ( 4 )   100753 - 100753   2020.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The molecular mechanisms underlying the 'seed and soil' theory are unknown. S100A8/A9 (a heterodimer complex of S100A8 and S100A9 proteins that exhibits a 'soil signal') is a ligand for Toll-like receptor 4, causing distant melanoma cells to approach the lung as a 'seeding' site. Unknown soil sensors for S100A8/A9 may exist, e.g., extracellular matrix metalloproteinase inducer, neuroplastin, activated leukocyte cell adhesion molecule, and melanoma cell adhesion molecule. We call these receptor proteins 'novel S100 soil sensor receptors (novel SSSRs).' Here we review and summarize a crucial role of the S100A8/A9-novel SSSRs' axis in cancer metastasis. The binding of S100A8/A9 to individual SSSRs is important in cancer metastasis via upregulations of the epithelial-mesenchymal transition, cellular motility, and cancer cell invasiveness, plus the formation of an inflammatory immune suppressive environment in metastatic organ(s). These metastatic cellular events are caused by the SSSR-featured signal transductions we identified that provide cancer cells a driving force for metastasis. To deprive cancer cells of these metastatic forces, we developed novel biologics that prevent the interaction of S100A8/A9 with SSSRs, followed by the efficient suppression of S100A8/A9-mediated lung-tropic metastasis in vivo.

    DOI: 10.1016/j.tranon.2020.100753

    PubMed

    researchmap

  • PLOD2 Is Essential to Functional Activation of Integrin β1 for Invasion/Metastasis in Head and Neck Squamous Cell Carcinomas. Reviewed International journal

    Yushi Ueki, Ken Saito, Hidekazu Iioka, Izumi Sakamoto, Yasuhiro Kanda, Masakiyo Sakaguchi, Arata Horii, Eisaku Kondo

    iScience   23 ( 2 )   100850 - 100850   2020.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Identifying the specific functional regulator of integrin family molecules in cancer cells is critical because they are directly involved in tumor invasion and metastasis. Here we report high expression of PLOD2 in oropharyngeal squamous cell carcinomas (SCCs) and its critical role as a stabilizer of integrin β1, enabling integrin β1 to initiate tumor invasion/metastasis. Integrin β1 stabilized by PLOD2-mediated hydroxylation was recruited to the plasma membrane, its functional site, and accelerated tumor cell motility, leading to tumor metastasis in vivo, whereas loss of PLOD2 expression abrogated it. In accordance with molecular analysis, examination of oropharyngeal SCC tissues from patients corroborated PLOD2 expression associated with integrin β1 at the invasive front of tumor nests. PLOD2 is thus implicated as the key regulator of integrin β1 that prominently regulates tumor invasion and metastasis, and it provides important clues engendering novel therapeutics for these intractable cancers.

    DOI: 10.1016/j.isci.2020.100850

    PubMed

    researchmap

  • Caco-2 cells monolayer as an in-vitro model for probiotic strain translocation

    Ni Nengah Dwi Fatmawati, Kazuyoshi Goto, I. Putu Bayu Mayura, Komang Ayu Nocianitri, Yan Ramona, Masakiyo Sakaguchi, Osamu Matsushita, I. Nengah Sujaya

    Bali Medical Journal   9 ( 1 )   137 - 142   2020

     More details

    Publishing type:Research paper (scientific journal)  

    Background: Caco-2 cells monolayer is one of in vitro models to evaluate the translocation capacity of Lactobacillus spp probiotic strains. The translocation is influenced by mucosa permeability of enterocytes as shown by increasing transepithelial resistance (TER) and formation of tight junction proteins. The pore size of the supported permeable membrane used in in vitro assay was one of the crucial factors in performing bacterial translocation assay. Almost no study has been conducted using Caco-2 cells monolayer grown on 8-μm pore size polycarbonate membrane for evaluating probiotics translocation. Therefore this study aimed to determine whether the Caco-2 cells monolayer model was suitable as an in vitro translocation model. Methods: Caco-2 cells monolayer was seeded onto 8-μm collagen-coated polycarbonate membrane insert Transwell®. Differentiation of Caco-2 cells was detected by measuring the TER, while the ZO-1 protein (the tight junction proteins) was detected by immunofluorescence. H2 O2 was used as a tight junction disruptive agent. Data were analyzed using SPSS version 23 software to compare the mean of TER measurement between untreated and H2 O2-treated Caco-2 cells monolayer. Results: The result showed that the TER of Caco-2 cells monolayer was gradually increasing until day 14, reaching more than 800 ohm.cm2. Furthermore, the ZO-1 protein was successfully detected, indicated the tight junction formation. TER value of H2 O2-treated cells showed significantly lower than that of untreated cells (P<0.05), indicating a disturbance of cells monolayer integrity. Lactobacillus rhamnosus FBB81 was used for validating the translocation. There was no translocation observed; however, translocation was observed in H2 O2-treated cells. Conclusion: Altogether suggests that Caco-2 cells grown on 8 μm-pore size permeable filters could be considered as a suitable in vitro model for probiotics strains translocation.

    DOI: 10.15562/bmj.v9i1.1633

    Scopus

    researchmap

  • PODXL1 promotes metastasis of the pancreatic ductal adenocarcinoma by activating the C5aR/C5a axis from the tumor microenvironment. Reviewed International journal

    Ken Saito, Hidekazu Iioka, Satoshi Maruyama, I Wayan Sumardika, Masakiyo Sakaguchi, Eisaku Kondo

    Neoplasia (New York, N.Y.)   21 ( 12 )   1121 - 1132   2019.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Pancreatic invasive ductal adenocarcinoma (PDAC) is a representative intractable malignancy under the current cancer therapies, and is considered a scirrhous carcinoma because it develops dense stroma. Both PODXL1, a member of CD34 family molecules, and C5aR, a critical cell motility inducer, have gained recent attention, as their expression was reported to correlate with poor prognosis for patients with diverse origins including PDAC; however, previous studies reported independently on their respective biological significance. Here we demonstrate that PODXL1 is essential for metastasis of PDAC cells through its specific interaction with C5aR. In vitro assay demonstrated that PODXL1 bound to C5aR, which stabilized C5aR protein and recruited it to cancer cell plasma membranes to receive C5a, an inflammatory chemoattractant factor. PODXL1 knockout in PDAC cells abrogated their metastatic property in vivo, emulating the liver metastatic mouse model treated with anti-C5a neutralizing antibody. In molecular studies, PODXL1 triggered EMT on PDAC cells in response to stimulation by C5a, corroborating PODXL1 involvement in PDAC cellular invasive properties via specific interaction with the C5aR/C5a axis. Confirming the molecular assays, histological examination showed coexpression of PODXL1 and C5aR at the invasive front of primary cancer nests as well as in liver metastatic foci of PDAC both in the mouse metastasis model and patient tissues. Hence, the novel direct interaction between PODXL1 and the C5aR/C5a axis may provide a better integrated understanding of PDAC biological characteristics including its tumor microenvironment factors.

    DOI: 10.1016/j.neo.2019.09.003

    PubMed

    researchmap

  • Crumbs3 promotes metastasis of colon cancer by regulating focal adhesion components Reviewed

    Iioka Hidekazu, Saito Ken, Sakaguchi Masakiyo, Morii Eiichi, Kondo Eisaku

    CANCER SCIENCE   145 ( 10 )   2740 - 2753   2019.11

  • Crumbs3 is a critical factor that regulates invasion and metastasis of colon adenocarcinoma via the specific interaction with FGFR1. Reviewed International journal

    Hidekazu Iioka, Ken Saito, Masakiyo Sakaguchi, Taro Tachibana, Keiichi Homma, Eisaku Kondo

    International journal of cancer   145 ( 10 )   2740 - 2753   2019.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Epithelial cell polarity regulator Crumbs3 (Crb3), a mammalian homolog within the Drosophila Crb gene family, was initially identified as an essential embryonic development factor. It is recently implicated in tumor suppression, though its specific functions are controversial. We here demonstrate that Crb3 strongly promotes tumor invasion and metastasis of human colon adenocarcinoma cells. Crb3 centrality to tumor migration was supported by strong expression at invasive front and metastatic foci of colonic adenocarcinoma of the patient tissues. Accordingly, two different Crb3-knockout (KO) lines, Crb3-KO (Crb3 -/-) DLD-1 and Crb3-KO WiDr from human colonic adenocarcinomas, were generated by the CRISPR-Cas9 system. Crb3-KO DLD-1 cells exhibited loss of cellular mobility in vitro and dramatic suppression of liver metastases in vivo in contrast to the wild type of DLD-1. Unlike DLD-1, Crb3-KO WiDr mobility and metastasis were unaffected, which were similar to wild-type WiDr. Proteome analysis of Crb3-coimmunopreciptated proteins identified different respective fibroblast growth factor receptor (FGFR) isotypes specifically bound to Crb3 isoform a through their intracellular domain. In DLD-1, Crb3 showed membranous localization of FGFR1 leading to its functional activation, whereas Crb3 bound to cytoplasmic FGFR4 in WiDr without FGFR1 expression, leading to cellular growth. Correlative expression between Crb3 and FGFR1 was consistently detected in primary and metastatic colorectal cancer patient tissues. Taking these together, Crb3 critically accelerates cell migration, namely invasion and metastasis of human colon cancers, through specific interaction to FGFR1 on colon cancer cells.

    DOI: 10.1002/ijc.32336

    PubMed

    researchmap

  • S100A11は筋浸潤性膀胱癌細胞と線維芽細胞のクロストークに関与し腫瘍進行に寄与する(S100A11 contributes to tumor progression with cross talking between muscle invasive bladder cancer cells and fibroblasts)

    光井 洋介, 定平 卓也, 渡部 昌実, 丸山 雄樹, 荒木 元朗, 渡邉 豊彦, 阪口 政清, 那須 保友

    西日本泌尿器科   81 ( 増刊 )   138 - 138   2019.10

     More details

    Language:English   Publisher:西日本泌尿器科学会  

    researchmap

  • Upregulation of Mobility in Pancreatic Cancer Cells by Secreted S100A11 Through Activation of Surrounding Fibroblasts. Reviewed International journal

    Yosuke Mitsui, Nahoko Tomonobu, Masami Watanabe, Rie Kinoshita, I Wayan Sumardika, Chen Youyi, Hitoshi Murata, Ken-Ichi Yamamoto, Takuya Sadahira, Acosta Gonzalez Herik Rodrigo, Hitoshi Takamatsu, Kota Araki, Akira Yamauchi, Masahiro Yamamura, Hideyo Fujiwara, Yusuke Inoue, Junichiro Futami, Ken Saito, Hidekazu Iioka, Eisaku Kondo, Masahiro Nishibori, Shinichi Toyooka, Yasuhiko Yamamoto, Yasutomo Nasu, Masakiyo Sakaguchi

    Oncology research   27 ( 8 )   945 - 956   2019.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    S100A11, a member of the S100 family of proteins, is actively secreted from pancreatic ductal adenocarcinoma (PDAC) cells. However, the role of the extracellular S100A11 in PDAC progression remains unclear. In the present study, we investigated the extracellular role of S100A11 in crosstalking between PDAC cells and surrounding fibroblasts in PDAC progression. An abundant S100A11 secreted from pancreatic cancer cells stimulated neighboring fibroblasts through receptor for advanced glycation end products (RAGE) upon S100A11 binding and was followed by not only an enhanced cancer cell motility in vitro but also an increased number of the PDAC-derived circulating tumor cells (CTCs) in vivo. Mechanistic investigation of RAGE downstream in fibroblasts revealed a novel contribution of a mitogen-activated protein kinase kinase kinase (MAPKKK), tumor progression locus 2 (TPL2), which is required for positive regulation of PDAC cell motility through induction of cyclooxygenase 2 (COX2) and its catalyzed production of prostaglandin E2 (PGE2), a strong chemoattractive fatty acid. The extracellularly released PGE2 from fibroblasts was required for the rise in cellular migration as well as infiltration of their adjacent PDAC cells in a coculture setting. Taken together, our data reveal a novel role of the secretory S100A11 in PDAC disseminative progression through activation of surrounding fibroblasts triggered by the S100A11-RAGE-TPL2-COX2 pathway. The findings of this study will contribute to the establishment of a novel therapeutic antidote to PDACs that are difficult to treat by regulating cancer-associated fibroblasts (CAFs) through targeting the identified pathway.

    DOI: 10.3727/096504019X15555408784978

    PubMed

    researchmap

  • Newly developed anti-S100A8/A9 monoclonal antibody efficiently prevents lung tropic cancer metastasis. Reviewed International journal

    Rie Kinoshita, Hiroki Sato, Akira Yamauchi, Yuta Takahashi, Yusuke Inoue, I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Kota Araki, Kazuhiko Shien, Shuta Tomida, Hidejiro Torigoe, Kei Namba, Eisuke Kurihara, Yusuke Ogoshi, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, Hiromasa Yamamoto, Junichi Soh, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    International journal of cancer   145 ( 2 )   569 - 575   2019.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The metastatic dissemination of cancer cells to remote areas of the body is the most problematic aspect in cancer patients. Among cancers, melanomas are notoriously difficult to treat due to their significantly high metastatic potential even during early stages. Hence, the establishment of advanced therapeutic approaches to regulate metastasis is required to overcome the melanoma disease. An accumulating mass of evidence has indicated a critical role of extracellular S100A8/A9 in melanoma distant metastasis. Lung S100A8/A9 is induced by melanoma cells from distant organs and it attracts these cells to its enriched lung environment since melanoma cells possess several receptors that sense the S100A8/A9 ligand. We hence aimed to develop a neutralizing antibody against S100A8/A9 that would efficiently block melanoma lung metastasis. Our protocol provided us with one prominent antibody, Ab45 that efficiently suppressed not only S100A8/A9-mediated melanoma mobility but also lung tropic melanoma metastasis in a mouse model. This prompted us to make chimeric Ab45, a chimera antibody consisting of mouse Ab45-Fab and human IgG2-Fc. Chimeric Ab45 also showed significant inhibition of the lung metastasis of melanoma. From these results, we have high hopes that the newly produced antibody will become a potential biological tool to block melanoma metastasis in future clinical settings.

    DOI: 10.1002/ijc.31982

    PubMed

    researchmap

  • Newly developed anti-S100A8/A9 monoclonal antibody efficiently prevents lung tropic cancer metastasis. Reviewed International journal

    Rie Kinoshita, Hiroki Sato, Akira Yamauchi, Yuta Takahashi, Yusuke Inoue, I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Kota Araki, Kazuhiko Shien, Shuta Tomida, Hidejiro Torigoe, Kei Namba, Eisuke Kurihara, Yusuke Ogoshi, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, Hiromasa Yamamoto, Junichi Soh, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    International journal of cancer   145 ( 2 )   569 - 575   2019.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The metastatic dissemination of cancer cells to remote areas of the body is the most problematic aspect in cancer patients. Among cancers, melanomas are notoriously difficult to treat due to their significantly high metastatic potential even during early stages. Hence, the establishment of advanced therapeutic approaches to regulate metastasis is required to overcome the melanoma disease. An accumulating mass of evidence has indicated a critical role of extracellular S100A8/A9 in melanoma distant metastasis. Lung S100A8/A9 is induced by melanoma cells from distant organs and it attracts these cells to its enriched lung environment since melanoma cells possess several receptors that sense the S100A8/A9 ligand. We hence aimed to develop a neutralizing antibody against S100A8/A9 that would efficiently block melanoma lung metastasis. Our protocol provided us with one prominent antibody, Ab45 that efficiently suppressed not only S100A8/A9-mediated melanoma mobility but also lung tropic melanoma metastasis in a mouse model. This prompted us to make chimeric Ab45, a chimera antibody consisting of mouse Ab45-Fab and human IgG2-Fc. Chimeric Ab45 also showed significant inhibition of the lung metastasis of melanoma. From these results, we have high hopes that the newly produced antibody will become a potential biological tool to block melanoma metastasis in future clinical settings.

    DOI: 10.1002/ijc.31982

    PubMed

    researchmap

  • Convenient methodology for extraction and subsequent selective propagation of mouse melanocytes in culture from adult mouse skin tissue. Reviewed International journal

    Nahoko Tomonobu, Rie Kinoshita, I Wayan Sumardika, Youyi Chen, Yusuke Inoue, Akira Yamauchi, Ken-Ichi Yamamoto, Hitoshi Murata, Masakiyo Sakaguchi

    Biochemistry and biophysics reports   18   100619 - 100619   2019.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Mouse melanoma B16-BL6 cells are useful cells for cancer metastatic studies. To understand the metastatic principle at molecular levels, it is necessary to carry out experiments in which cancer cells and their normal counterparts are compared. However, unlike normal human melanocytes, preparation of normal mouse melanocytes is quite difficult due to the lack of marketing and insufficient information on an established protocol for primary culture of mouse melanocytes. In this study, we aimed to establish a convenient method for primary culture of mouse melanocytes on the basis of the protocol for human melanocytes. The main obstacles to preparing pure mouse melanocytes are how to digest mouse skin tissue and how to reduce the contamination of keratinocytes and fibroblasts. The obstacles were overcome by collagenase digestion for skin specimens, short time trypsinization for separating melanocytes and keratinocytes, and use of 12-O-Tetradecanoylphorbol 13-acetate (TPA) and cholera toxin in the culture medium. These supplements act to prevent the proliferation of keratinocytes and fibroblasts, respectively. The convenient procedure enabled us to prepare a pure culture of normal mouse melanocytes. Using enriched normal mouse melanocytes and cancerous B16-BL6 cells, we compared the expression levels of melanoma cell adhesion molecule (MCAM), an important membrane protein for melanoma metastasis, in the cells. The results showed markedly higher expression of MCAM in B16-BL6 cells than in normal mouse melanocytes.

    DOI: 10.1016/j.bbrep.2019.100619

    PubMed

    researchmap

  • Critical role of the MCAM-ETV4 axis triggered by extracellular S100A8/A9 in breast cancer aggressiveness. Reviewed International journal

    Youyi Chen, I Wayan Sumardika, Nahoko Tomonobu, Rie Kinoshita, Yusuke Inoue, Hidekazu Iioka, Yosuke Mitsui, Ken Saito, I Made Winarsa Ruma, Hiroki Sato, Akira Yamauchi, Hitoshi Murata, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiromasa Yamamoto, Junichi Soh, Junichiro Futami, Miyoko Kubo, Endy Widya Putranto, Takashi Murakami, Ming Liu, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    Neoplasia (New York, N.Y.)   21 ( 7 )   627 - 640   2019.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Metastatic breast cancer is the leading cause of cancer-associated death in women. The progression of this fatal disease is associated with inflammatory responses that promote cancer cell growth and dissemination, eventually leading to a reduction of overall survival. However, the mechanism(s) of the inflammation-boosted cancer progression remains unclear. In this study, we found for the first time that an extracellular cytokine, S100A8/A9, accelerates breast cancer growth and metastasis upon binding to a cell surface receptor, melanoma cell adhesion molecule (MCAM). Our molecular analyses revealed an important role of ETS translocation variant 4 (ETV4), which is significantly activated in the region downstream of MCAM upon S100A8/A9 stimulation, in breast cancer progression in vitro as well as in vivo. The MCAM-mediated activation of ETV4 induced a mobile phenotype called epithelial-mesenchymal transition (EMT) in cells, since we found that ETV4 transcriptionally upregulates ZEB1, a strong EMT inducer, at a very high level. In contrast, downregulation of either MCAM or ETV4 repressed EMT, resulting in greatly weakened tumor growth and lung metastasis. Overall, our results revealed that ETV4 is a novel transcription factor regulated by the S100A8/A9-MCAM axis, which leads to EMT through ZEB1 and thereby to metastasis in breast cancer cells. Thus, therapeutic strategies based on our findings might improve patient outcomes.

    DOI: 10.1016/j.neo.2019.04.006

    PubMed

    researchmap

  • Melanoma cell adhesion molecule is the driving force behind the dissemination of melanoma upon S100A8/A9 binding in the original skin lesion. Reviewed International journal

    Youyi Chen, I Wayan Sumardika, Nahoko Tomonobu, I Made Winarsa Ruma, Rie Kinoshita, Eisaku Kondo, Yusuke Inoue, Hiroki Sato, Akira Yamauchi, Hitoshi Murata, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiromasa Yamamoto, Junichi Soh, Ming Liu, Junichiro Futami, Kaori Sasai, Hiroshi Katayama, Miyoko Kubo, Endy Widya Putranto, Toshihiko Hibino, Bei Sun, Masahiro Nishibori, Shinichi Toyooka, Masakiyo Sakaguchi

    Cancer letters   452   178 - 190   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Since metastasis accounts for the majority of cancer-associated deaths, studies on the mechanisms of metastasis are needed to establish innovative strategies for cancer treatment. We previously reported that melanoma cell adhesion molecule (MCAM) functions as a critical receptor for S100A8/A9, and binding of S100A8/A9 to MCAM results in the migration of melanoma cells to lung tissue. However, the critical role of MCAM in the original melanoma skin lesion is still not clear. In this study, we aimed to determine the importance of the S100A8/A9-MCAM axis in melanoma dissemination in a skin lesion as a critical early step for metastasis. Mechanistic studies revealed the downstream signaling of MCAM that signaled the induction of metastasis. S100A8/A9-MCAM binding activates mitogen-activated protein kinase kinase kinase 8 (MAP3K8), also termed TPL2, leading to strong activation of the transcription factor ETV4 and subsequent induction of matrix metalloproteinase-25 (MMP25), and finally to induction of melanoma lung tropic metastasis. Collectively, our results demonstrate a crucial role of the S100A8/A9-MCAM signaling axis in metastatic onset of melanoma cells and indicate that strategies targeting the identified pathway may be useful for the establishment of innovative anti-cancer therapies.

    DOI: 10.1016/j.canlet.2019.03.023

    PubMed

    researchmap

  • Extracellular S100A11 Plays a Critical Role in Spread of the Fibroblast Population in Pancreatic Cancers. Reviewed International journal

    Hitoshi Takamatsu, Ken-Ichi Yamamoto, Nahoko Tomonobu, Hitoshi Murata, Yusuke Inoue, Akira Yamauchi, I Wayan Sumardika, Youyi Chen, Rie Kinoshita, Masahiro Yamamura, Hideyo Fujiwara, Yosuke Mitsui, Kota Araki, Junichiro Futami, Ken Saito, Hidekazu Iioka, I Made Winarsa Ruma, Endy Widya Putranto, Masahiro Nishibori, Eisaku Kondo, Yasuhiko Yamamoto, Shinichi Toyooka, Masakiyo Sakaguchi

    Oncology research   27 ( 6 )   713 - 727   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    The fertile stroma in pancreatic ductal adenocarcinomas (PDACs) has been suspected to greatly contribute to PDAC progression. Since the main cell constituents of the stroma are fibroblasts, there is crosstalking(s) between PDAC cells and surrounding fibroblasts in the stroma, which induces a fibroblast proliferation burst. We have reported that several malignant cancer cells including PDAC cells secrete a pronounced level of S100A11, which in turn stimulates proliferation of cancer cells via the receptor for advanced glycation end products (RAGE) in an autocrine manner. Owing to the RAGE+ expression in fibroblasts, the extracellular abundant S100A11 will affect adjacent fibroblasts. In this study, we investigated the significance of the paracrine axis of S100A11-RAGE in fibroblasts for their proliferation activity. In in vitro settings, extracellular S100A11 induced upregulation of fibroblast proliferation. Our mechanistic studies revealed that the induction is through RAGE-MyD88-mTOR-p70 S6 kinase upon S100A11 stimulation. The paracrine effect on fibroblasts is linked mainly to triggering growth but not cellular motility. Thus, the identified pathway might become a potential therapeutic target to suppress PDAC progression through preventing PDAC-associated fibroblast proliferation.

    DOI: 10.3727/096504018X15433161908259

    PubMed

    researchmap

  • exSSSRs (extracellular S100 soil sensor receptors)-Fc fusion proteins work as prominent decoys to S100A8/A9-induced lung tropic cancer metastasis. Reviewed International journal

    Rie Kinoshita, Hiroki Sato, Akira Yamauchi, Yuta Takahashi, Yusuke Inoue, I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Kota Araki, Kazuhiko Shien, Shuta Tomida, Hidejiro Torigoe, Kei Namba, Eisuke Kurihara, Yusuke Ogoshi, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, Hiromasa Yamamoto, Junichi Soh, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    International journal of cancer   144 ( 12 )   3138 - 3145   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Within the "seed and soil" theory of organ tropic cancer metastasis is a growing compilation of evidence that S100A8/A9 functions as a soil signal that attracts cancer cells to certain organs, which prove beneficial to their growth. S100A8/A9-sensing receptors including Toll-like receptor 4 (TLR4), advanced glycation end products (RAGE), and also important receptors we recently succeeded in identifying (EMMPRIN, NPTNβ, MCAM, and ALCAM) have the potential to become promising therapeutic targets. In our study, we prepared extracellular regions of these novel molecules and fused them to human IgG2-Fc to extend half-life expectancy, and we evaluated the anti-metastatic effects of the purified decoy proteins on metastatic cancer cells. The purified proteins markedly suppressed S100A8/A9-mediated lung tropic cancer metastasis. We hence expect that our novel biologics may become a prominent medicine to prevent cancer metastasis in clinical settings through cutting the linkage between "seed and soil".

    DOI: 10.1002/ijc.31945

    PubMed

    researchmap

  • Neuroplastin-β mediates S100A8/A9-induced lung cancer disseminative progression. Reviewed International journal

    I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Rie Kinoshita, I Made Winarsa Ruma, Hiroki Sato, Eisaku Kondo, Yusuke Inoue, Akira Yamauchi, Hitoshi Murata, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiromasa Yamamoto, Junichi Soh, Junichiro Futami, Endy Widya Putranto, Toshihiko Hibino, Masahiro Nishibori, Shinichi Toyooka, Masakiyo Sakaguchi

    Molecular carcinogenesis   58 ( 6 )   980 - 995   2019.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Compiling evidence indicates an unusual role of extracellular S100A8/A9 in cancer metastasis. S100A8/A9 secreted from either cancer cells or normal cells including epithelial and inflammatory cells stimulates cancer cells through S100A8/A9 sensor receptors in an autocrine or paracrine manner, leading to cancer cell metastatic progression. We previously reported a novel S100A8/A9 receptor, neuroplastin-β (NPTNβ), which plays a critical role in atopic dermatitis when it is highly activated in keratinocytes by an excess amount of extracellular S100A8/A9 in the inflammatory skin lesion. Interestingly, our expression profiling of NPTNβ showed significantly high expression levels in lung cancer cell lines in a consistent manner. We hence aimed to determine the significance of NPTNβ as an S100A8/A9 receptor in lung cancer. Our results showed that NPTNβ has strong ability to induce cancer-related cellular events, including anchorage-independent growth, motility and invasiveness, in lung cancer cells in response to extracellular S100A8/A9, eventually leading to the expression of a cancer disseminative phenotype in lung tissue in vivo. Mechanistic investigation revealed that binding of S100A8/A9 to NPTNβ mediates activation of NFIA and NFIB and following SPDEF transcription factors through orchestrated upstream signals from TRAF2 and RAS, which is linked to anchorage-independent growth, motility and invasiveness. Overall, our results indicate the importance of the S100A8/A9-NPTNβ axis in lung cancer disseminative progression and reveal a pivotal role of its newly identified downstream signaling, TRAF2/RAS-NFIA/NFIB-SPDEF, in linking to the aggressive development of lung cancers.

    DOI: 10.1002/mc.22987

    PubMed

    researchmap

  • Histidine-rich glycoprotein augments natural killer cell function by modulating PD-1 expression via CLEC-1B. Reviewed International journal

    Nishimura Y, Wake H, Teshigawara K, Wang D, Sakaguchi M, Otsuka F, Nishibori M

    Pharmacol Res Perspect.   7 ( 3 )   e00481   2019.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Augmentation of natural killer (NK) cell cytotoxicity is one of the greatest challenges for cancer immunotherapy. Although histidine-rich glycoprotein (HRG), a 75-kDa glycoprotein with various immunomodulatory activities, reportedly elicits antitumor immunity, its effect on NK cell cytotoxicity is unclear. We assessed NK cell cytotoxicity against K562 cells. We also measured concentrations of cytokines and granzyme B in the cell supernatant. The proportion of CD56bright NK cells and NK cell surface PD-1 expression was assessed with flow cytometry. The neutralizing effects of anti-C-type lectin-like receptor (CLEC) 1B against HRG were also measured. NK cell morphological changes were analyzed via confocal microscopy. HRG significantly increased NK cell cytotoxicity against K562 cell lines. HRG also increased the release of granzyme B and the proportion of CD56bright NK cells. Further, HRG was able to decrease NK cell surface PD-1 expression. The effects of HRG on NK cells were reversed with anti-CLEC-1B antibodies. Additionally, we confirmed NK cell nuclear morphology and F-actin distribution, which are involved in the regulation of cytotoxic granule secretion. Because both PD-1 and CLEC-1B are associated with prognosis during malignancy, HRG incorporates these molecules to exert the antitumor immunity role. These facts indicate the potential of HRG to be a new target for cancer immunotherapy.

    DOI: 10.1002/prp2.481.

    PubMed

    researchmap

  • Hepatocyte nuclear factor 4α regulates megalin expression in proximal tubular cells. Reviewed International journal

    Shota Sasaki, Ayami Hara, Masakiyo Sakaguchi, Masaomi Nangaku, Yusuke Inoue

    Biochemistry and biophysics reports   17   87 - 92   2019.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Hepatocyte nuclear factor 4α (HNF4α) is a member of the nuclear receptor superfamily and upregulates expression of many genes in the liver, pancreas, small intestine, and colon. HNF4α is also highly expressed in proximal tubular epithelial cells (PTECs) in kidney. PTECs reabsorb various substances through transporters, ion channels, and receptors, but the target genes for HNF4α in PTECs have not been investigated in detail. In the present study, we aimed to identify novel HNF4α target genes that are highly expressed in PTECs. Expression of many solute carrier transporter genes was upregulated by HNF4α in human PTEC-derived HK-2 cells. Notably, expression of megalin (LRP2), an endocytic receptor of various molecules involved in development and progression of chronic kidney disease (CKD), was strongly induced by HNF4α, and the transactivation potential of the megalin promoter was dependent on HNF4α expression. Moreover, HNF4α was found to directly bind to an HNF4α binding site near the transcription start site in the megalin gene. These results indicate that HNF4α plays an important role in maintaining reabsorption and metabolism in PTECs by positive regulation of several solute carrier transporter and megalin genes at the transcriptional level.

    DOI: 10.1016/j.bbrep.2018.11.010

    PubMed

    researchmap

  • Anti-tumor effect of neratinib against lung cancer cells harboring HER2 oncogene alterations. Reviewed International journal

    Yusuke Ogoshi, Kazuhiko Shien, Takahiro Yoshioka, Hidejiro Torigoe, Hiroki Sato, Masakiyo Sakaguchi, Shuta Tomida, Kei Namba, Eisuke Kurihara, Yuta Takahashi, Ken Suzawa, Hiromasa Yamamoto, Junichi Soh, Shinichi Toyooka

    Oncology letters   17 ( 3 )   2729 - 2736   2019.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Human epidermal growth factor receptor 2 (HER2) is a member of the ErbB family of receptor tyrosine kinases. Numerous studies have reported the amplification and overexpression of HER2 in several types of cancer, including non-small cell lung cancer (NSCLC). However, the benefits of HER2-targeted therapy have not been fully established. In the present study, the anti-tumor effect of neratinib, an irreversible pan-HER tyrosine kinase inhibitor (TKI), against NSCLC cells harboring HER2 alterations was investigated. The sensitivity of normal bronchial epithelial cells (BEAS-2B) ectopically overexpressing wild-type or mutant HER2 to neratinib was assessed. Furthermore, the anti-tumor activity of neratinib in several NSCLC cell lines harboring HER2 alterations was determined in vitro and in vivo, and the association between their genetic alterations and sensitivity to neratinib treatment was investigated. BEAS-2B cells ectopically overexpressing wild-type HER2 or mutants (A775insYVMA, G776VC, G776LC, P780insGSP, V659E, G660D and S310F) exhibited constitutive autophosphorylation of HER2, as determined by western blotting. While these BEAS-2B cells were sensitive to neratinib, they were insensitive to erlotinib, a first-generation epidermal growth factor receptor-TKI. Neratinib also exerted anti-proliferative effects on HER2-altered (H2170, Calu-3 and H1781) NSCLC cell lines. Neratinib was also demonstrated to exert strong tumor growth inhibitory activity in mouse xenograft models using HER2-altered lung cancer cells. The results of the present study strongly suggest that neratinib has potential as a promising therapeutic option for the treatment of HER2-altered NSCLC.

    DOI: 10.3892/ol.2019.9908

    PubMed

    researchmap

  • c-Jun N-terminal kinase (JNK)-mediated phosphorylation of SARM1 regulates NAD+ cleavage activity to inhibit mitochondrial respiration. Reviewed International journal

    Hitoshi Murata, Cho Cho Khine, Akane Nishikawa, Ken-Ichi Yamamoto, Rie Kinoshita, Masakiyo Sakaguchi

    The Journal of biological chemistry   293 ( 49 )   18933 - 18943   2018.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Mitochondrial dysfunction is a key pathological feature of many different types of neurodegenerative disease. Sterile alpha and Toll/interleukin receptor motif-containing protein 1 (SARM1) has been attracting much attention as an important molecule for inducing axonal degeneration and neuronal cell death by causing loss of NAD (NADH). However, it has remained unclear what exactly regulates the SARM1 activity. Here, we report that NAD+ cleavage activity of SARM1 is regulated by its own phosphorylation at serine 548. The phosphorylation of SARM1 was mediated by c-jun N-terminal kinase (JNK) under oxidative stress conditions, resulting in inhibition of mitochondrial respiration concomitant with enhanced activity of NAD+ cleavage. Nonphosphorylatable mutation of Ser-548 or treatment with a JNK inhibitor decreased SARM1 activity. Furthermore, neuronal cells derived from a familial Parkinson's disease (PD) patient showed a congenitally increased level of SARM1 phosphorylation compared with that in neuronal cells from a healthy person and were highly sensitive to oxidative stress. These results indicate that JNK-mediated phosphorylation of SARM1 at Ser-548 is a regulator of SARM1 leading to inhibition of mitochondrial respiration. These findings suggest that an abnormal regulation of SARM1 phosphorylation is involved in the pathogenesis of Parkinson's disease and possibly other neurodegenerative diseases.

    DOI: 10.1074/jbc.RA118.004578

    PubMed

    researchmap

  • JNK-mediated phosphorylation of SARM1 regulates NAD(+) cleavage activity to inhibit mitochondrial respiration. Reviewed

    Murata H, Khine CC, Nishikawa A, Yamamoto KI, Kinoshita R, Sakaguchi M

    J Biol Chem   2018.10

  • Combined inhibition of MEK and PI3K pathways overcomes acquired resistance to EGFR-TKIs in non-small cell lung cancer. Reviewed International journal

    Hiroki Sato, Hiromasa Yamamoto, Masakiyo Sakaguchi, Kazuhiko Shien, Shuta Tomida, Tadahiko Shien, Hirokuni Ikeda, Minami Hatono, Hidejiro Torigoe, Kei Namba, Takahiro Yoshioka, Eisuke Kurihara, Yusuke Ogoshi, Yuta Takahashi, Junichi Soh, Shinichi Toyooka

    Cancer science   109 ( 10 )   3183 - 3196   2018.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Compensatory activation of the signal transduction pathways is one of the major obstacles for the targeted therapy of non-small cell lung cancer (NSCLC). Herein, we present the therapeutic strategy of combined targeted therapy against the MEK and phosphoinositide-3 kinase (PI3K) pathways for acquired resistance to epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) in NSCLC. We investigated the efficacy of combined trametinib plus taselisib therapy using experimentally established EGFR-TKI-resistant NSCLC cell lines. The results showed that the feedback loop between MEK/ERK and PI3K/AKT pathways had developed in several resistant cell lines, which caused the resistance to single-agent treatment with either inhibitor alone. Meanwhile, the combined therapy successfully regulated the compensatory activation of the key intracellular signals and synergistically inhibited the cell growth of those cells in vitro and in vivo. The resistance mechanisms for which the dual kinase inhibitor therapy proved effective included (MET) mesenchymal-epithelial transition factor amplification, induction of epithelial-to-mesenchymal transition (EMT) and EGFR T790M mutation. In further analysis, the combination therapy induced the phosphorylation of p38 MAPK signaling, leading to the activation of apoptosis cascade. Additionally, long-term treatment with the combination therapy induced the conversion from EMT to mesenchymal-to-epithelial transition in the resistant cell line harboring EMT features, restoring the sensitivity to EGFR-TKI. In conclusion, our results indicate that the combined therapy using MEK and PI3K inhibitors is a potent therapeutic strategy for NSCLC with the acquired resistance to EGFR-TKIs.

    DOI: 10.1111/cas.13763

    PubMed

    researchmap

  • Induction of hepatic metabolic functions by a novel variant of hepatocyte nuclear factor 4γ. Reviewed

    Sasaki S, Urabe M, Maeda T, Suzuki J, Irie R, Suzuki M, Tomaru Y, Sakaguchi M, Gonzalez FJ, Inoue Y

    Molecular and cellular biology   38 ( 24 )   e00213 - e00218   2018.9

  • EGFR変異陽性肺癌におけるLRIG1の抗腫瘍効果(Tumor-suppressive effect of LRIG1 in non-small cell lung cancer harboring mutant EGFR)

    鳥越 英次郎, 山本 寛斉, 阪口 政清, 難波 圭, 佐藤 博紀, 枝園 和彦, 諏澤 憲, 宗 淳一, 冨田 秀太, 佃 和憲, 三好 新一郎, 豊岡 伸一

    日本癌学会総会記事   77回   940 - 940   2018.9

     More details

    Language:English   Publisher:(一社)日本癌学会  

    researchmap

  • Embigin Promotes Prostate Cancer Progression by S100A4-Dependent and-Independent Mechanisms. Reviewed International journal

    I Made Winarsa Ruma, Rie Kinoshita, Nahoko Tomonobu, Yusuke Inoue, Eisaku Kondo, Akira Yamauchi, Hiroki Sato, I Wayan Sumardika, Youyi Chen, Ken-Ichi Yamamoto, Hitoshi Murata, Shinichi Toyooka, Masahiro Nishibori, Masakiyo Sakaguchi

    Cancers   10 ( 7 )   2018.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Embigin, a transmembrane glycoprotein belonging to the immunoglobulin superfamily, is involved in prostate and mammary gland development. As embigin's roles in cancer remain elusive, we studied its biological functions and interaction with extracellular S100A4 in prostate cancer progression. We found by a pull-down assay that embigin is a novel receptor for S100A4, which is one of the vital cancer microenvironment milleu. Binding of extracellular S100A4 to embigin mediates prostate cancer progression by inhibition of AMPK activity, activation of NF-κB, MMP9 and mTORC1 signaling, and inhibition of autophagy, which increase prostate cancer cell motility. We also found that embigin promotes prostate cancer growth, spheroid- and colony-forming ability, and survival upon chemotherapy independently of S100A4. An in vivo growth mouse model confirmed the importance of embigin and its cytoplasmic tail in mediating prostate tumor growth. Moreover, embigin and p21WAF1 can be used to predict survival of prostate cancer patients. Our results demonstrated for the first time that the S100A4-embigin/AMPK/mTORC1/p21WAF1 and NF-κB/MMP9 axis is a vital oncogenic molecular cascade for prostate cancer progression. We proposed that embigin and p21WAF1 could be used as prognostic biomarkers and a strategy to inhibit S100A4-embigin binding could be a therapeutic approach for prostate cancer patients.

    DOI: 10.3390/cancers10070239

    PubMed

    researchmap

  • 膵がん進展に導く膵がん細胞 間質線維芽細胞クロストークを介在する分泌性S100A11 受容体RAGE連携の役割

    光井 洋介, 山本 健一, Sumardika I Wayan, 木下 理恵, 村田 等, 二見 淳一郎, 高松 仁, 山本 靖彦, 西堀 正洋, 豊岡 伸一, 渡部 昌実, 那須 保友, 阪口 政清

    日本がん免疫学会総会プログラム・抄録集   22回   156 - 156   2018.7

     More details

    Language:Japanese   Publisher:日本がん免疫学会  

    researchmap

  • 分泌性S100A11-受容体RAGEシグナルに着眼した膵がん間質増大のメカニズムの解明

    山本 健一, 高松 仁, 光井 洋介, 木下 理恵, 村田 等, 二見 淳一郎, 山本 靖彦, 西堀 正洋, 豊岡 伸一, 阪口 政清

    日本がん免疫学会総会プログラム・抄録集   22回   117 - 117   2018.7

     More details

    Language:Japanese   Publisher:日本がん免疫学会  

    researchmap

  • Tumor-suppressive effect of LRIG1, a negative regulator of ErbB, in non-small cell lung cancer harboring mutant EGFR. Reviewed International journal

    Hidejiro Torigoe, Hiromasa Yamamoto, Masakiyo Sakaguchi, Chen Youyi, Kei Namba, Hiroki Sato, Kazuhiko Shien, Junichi Soh, Ken Suzawa, Shuta Tomida, Kazunori Tsukuda, Shinichiro Miyoshi, Shinichi Toyooka

    Carcinogenesis   39 ( 5 )   719 - 727   2018.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Epidermal growth factor receptor (EGFR) is a member of the ErbB (HER) family that is known to play important roles in the pathogenesis of various human cancers. Mutations of the EGFR gene are commonly found as oncogenic driver mutations and have been targeted for treatment of non-small cell lung cancer (NSCLC). Leucine-rich repeat and immunoglobulin-like domain protein-1 (LRIG1) is a cell-surface protein that is known as a negative regulator of the ErbB (HER) family. In this study, we first confirmed that the expression levels of LRIG1 were much lower in NSCLC than in non-malignant cells or tissues. Next, we focused on the effect of LRIG1 in NSCLC. For this purpose, we established clones stably overexpressing LRIG1, using EGFR-mutant (HCC827, HCC4011 and NCI-H1975) and wild-type (A549) cells. Transfection of LRIG1 was associated with a decrease in the expression and phosphorylation levels of EGFR in the HCC827, HCC4011 and NCI-H1975 cells. It was also associated with strong suppression of the cell proliferative, invasive, migratory and tumorigenic potential of the HCC827 cells. On the other hand, no such effects were observed in the A549 cells. In addition, LRIG1 also downregulated the expression and phosphorylation levels of other tyrosine kinase receptors, such as HER2, HER3, MET and IGF-1R, and prevented the epithelial-to-mesenchymal transition induced by TGF-β in the HCC827 cells. These findings suggest that LRIG1 exerts important tumor-suppressive effects in EGFR-mutant NSCLC and has the potential to become a novel therapeutic target for EGFR-mutant NSCLC.

    DOI: 10.1093/carcin/bgy044

    Web of Science

    PubMed

    researchmap

  • Tumor necrosis factor- downregulates the REIC/Dkk-3 tumor suppressor gene in normal human skin keratinocytes Reviewed

    Ken Kataoka, Natsumi Maehara, Yuki Ayabe, Hitoshi Murata, Nam-Ho Huh, Masakiyo Sakaguchi

    Molecular Medicine Reports   17 ( 5 )   6661 - 6666   2018.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Spandidos Publications  

    Our preeious studies reeealed that REIC/Dkk-3 was expressed earious tissues, including skin keratinocytes. The aim of the present study was to identify the factors that regulate the expression of the dickkopf Wnt signaling pathway inhibitor 3 (REIC/Dkk-3) tumor suppressor gene in normal human skin keratinocytes (NHKs). Seeeral growth factors and cytokines that haee preeiously been reported to be ineoleed in the growth and differentiation of keratinocytes were screened as potential regulators. Western blot analysis was performed using protein from NHKs cultured with/without earious factors including the epidermal growth factor, tumor necrosis factor-, transforming growth factor-, interleukin (IL)-1F9, IL-6, IL-8 and Ca2+. The results indicated that only TNF- downregulated REIC/Dkk-3 expression in NHKs. Subsequently, TNF- was confirmed to reduce the expression leeels of REIC/Dkk-3 in mouse skin tissue and hair culture models. TNF-mediated downregulation of REIC/Dkk-3 expression in NHKs was abrogated by the addition of a TNF-specific antibody. In conclusion, the results indicate that TNF- downregulates REIC/Dkk-3 expression in normal skin keratinocytes.

    DOI: 10.3892/mmr.2018.8676

    Web of Science

    Scopus

    PubMed

    researchmap

  • Therapeutic strategies for afatinib-resistant lung cancer harboring HER2 alterations Reviewed

    Hidejiro Torigoe, Kazuhiko Shien, Tatsuaki Takeda, Takahiro Yoshioka, Kei Namba, Hiroki Sato, Ken Suzawa, Hiromasa Yamamoto, Junichi Soh, Masakiyo Sakaguchi, Shuta Tomida, Kazunori Tsukuda, Shinichiro Miyoshi, Shinichi Toyooka

    Cancer Science   109 ( 5 )   1493 - 1502   2018.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Blackwell Publishing Ltd  

    Human epidermal growth factor receptor 2 (HER2) plays an important role in the pathogenesis of various cancers. HER2 alterations have been suggested to be a therapeutic target in non-small-cell lung cancer (NSCLC), just as in breast and gastric cancers. We previously reported that the pan-HER inhibitor afatinib could be a useful therapeutic agent as HER2-targeted therapy for patients with NSCLC harboring HER2 alterations. However, acquired resistance to afatinib was observed in the clinical setting, similar to the case for other HER inhibitors. Thus, elucidation of the mechanisms underlying the development of acquired drug resistance and exploring means to overcome acquired drug resistance are important issues in the treatment of NSCLC. In this study, we experimentally established afatinib-resistant cell lines from NSCLC cell lines harboring HER2 alterations, and investigated the mechanisms underlying the acquisition of drug resistance. The established cell lines showed several unique afatinib-resistance mechanisms, including MET amplification, loss of HER2 amplification and gene expression, epithelial-to-mesenchymal transition (EMT) and acquisition of cancer stem cell (CSC)-like features. The afatinib-resistant cell lines showing MET amplification were sensitive to the combination of afatinib plus crizotinib (a MET inhibitor), both in vitro and in vivo. The resistant cell lines which showed EMT or had acquired CSC-like features remained sensitive to docetaxel, like the parental cells. These findings may provide clues to countering the resistance to afatinib in NSCLC patients with HER2 alterations.

    DOI: 10.1111/cas.13571

    Web of Science

    Scopus

    PubMed

    researchmap

  • β-1,3-Galactosyl-O-Glycosyl-Glycoprotein β-1,6-N-Acetylglucosaminyltransferase 3 Increases MCAM Stability, Which Enhances S100A8/A9-Mediated Cancer Motility. Reviewed International journal

    I Wayan Sumardika, Chen Youyi, Eisaku Kondo, Yusuke Inoue, I Made Winarsa Ruma, Hitoshi Murata, Rie Kinoshita, Ken-Ichi Yamamoto, Shuta Tomida, Kazuhiko Shien, Hiroki Sato, Akira Yamauchi, Junichiro Futami, Endy Widya Putranto, Toshihiko Hibino, Shinichi Toyooka, Masahiro Nishibori, Masakiyo Sakaguchi

    Oncology research   26 ( 3 )   431 - 444   2018.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    We previously identified novel S100A8/A9 receptors, extracellular matrix metalloproteinase inducer (EMMPRIN), melanoma cell adhesion molecule (MCAM), activated leukocyte cell adhesion molecule (ALCAM), and neuroplastin (NPTN) β, that are critically involved in S100A8/A9-mediated cancer metastasis and inflammation when expressed at high levels. However, little is known about the presence of any cancer-specific mechanism(s) that modifies these receptors, further inducing upregulation at protein levels without any transcriptional regulation. Expression levels of glycosyltransferase-encoding genes were examined by a PCR-based profiling array followed by confirmation with quantitative real-time PCR. Cell migration and invasion were assessed using a Boyden chamber. Western blotting was used to examine the protein level, and the RNA level was examined by Northern blotting. Immunohistochemistry was used to examine the expression pattern of β-1,3-galactosyl-O-glycosyl-glycoprotein β-1,6-N-acetylglucosaminyltransferase 3 (GCNT3) and MCAM in melanoma tissue. We found that GCNT3 is overexpressed in highly metastatic melanomas. Silencing and functional inhibition of GCNT3 greatly suppressed migration and invasion of melanoma cells, resulting in the loss of S100A8/A9 responsiveness. Among the novel S100A8/A9 receptors, GCNT3 favorably glycosylates the MCAM receptor, extending its half-life and leading to further elevation of S100A8/A9-mediated cellular motility in melanoma cells. GCNT3 expression is positively correlated to MCAM expression in patients with high-grade melanomas. Collectively, our results showed that GCNT3 is an upstream regulator of MCAM protein and indicate the possibility of a potential molecular target in melanoma therapeutics through abrogation of the S100A8/A9-MCAM axis.

    DOI: 10.3727/096504017X15031557924123

    Web of Science

    PubMed

    researchmap

  • Effects of Histidine-rich glycoprotein on erythrocyte aggregation and hemolysis: Implications for a role under septic conditions Reviewed

    Hui Zhong, Hidenori Wake, Keyue Liu, Yuan Gao, Kiyoshi Teshigawara, Masakiyo Sakaguchi, Shuji Mori, Masahiro Nishibori

    Journal of Pharmacological Sciences   136 ( 3 )   97 - 106   2018.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japanese Pharmacological Society  

    The apoptotic process of erythrocytes is known as eryptosis, and is characterized by phosphatidylserine (PS) expression on the outer membrane. PS-positive erythrocytes are increased in sepsis, and PS is believed to facilitate coagulation of erythrocytes and activate macrophages. However, the relationship between eryptosis and abnormal coagulation in sepsis is still not fully understood. Histidine-rich glycoprotein (HRG) inhibits immunothrombus formation by regulating neutrophils and vascular endothelial cells. In the present study, we subjected isolated erythrocytes to Zn2+ stimulation, which activated their aggregation and PS expression. We then determined the Zn2+ contents in septic lung and kidney tissues, and found that they were elevated, suggesting that eryptosis was enhanced in these tissues. Erythrocyte adhesion to endothelial cells was also significantly increased after Zn2+ stimulation, and this effect was inhibited by HRG. Finally, we examined HRG treatment in septic model mice, and found that HRG decreased hemolysis, possibly due to its ability to bind heme. Our study demonstrated a novel Zn2+-initiated aggregation/thrombus formation pathway. We also showed the regulatory role of HRG in this pathway, together with the ability of HRG to inhibit hemolysis under septic conditions. HRG supplementation might be a novel therapeutic strategy for inflammatory disorders, especially sepsis.

    DOI: 10.1016/j.jphs.2017.11.003

    Web of Science

    Scopus

    PubMed

    researchmap

  • Therapeutic potential of targeting S100A11 in malignant pleural mesothelioma Reviewed

    Hiroki Sato, Masakiyo Sakaguchi, Hiromasa Yamamoto, Shuta Tomida, Keisuke Aoe, Kazuhiko Shien, Takahiro Yoshioka, Kei Namba, Hidejiro Torigoe, Junichi Soh, Kazunori Tsukuda, Hiroyuki Tao, Kazunori Okabe, Shinichiro Miyoshi, Harvey I. Pass, Shinichi Toyooka

    Oncogenesis   7 ( 1 )   11   2018.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Nature Publishing Group  

    Malignant pleural mesothelioma (MPM) is an aggressive tumor with an unfavorable prognosis. The standard therapeutic approaches are limited to surgery, chemotherapy, and radiotherapy. Because the consequent clinical outcome is often unsatisfactory, a different approach in MPM treatment is required. S100A11, a Ca2+-binding small protein with two EF-hands, is frequently upregulated in various human cancers. Interestingly, it has been found that intracellular and extracellular S100A11 have different functions in cell viability. In this study, we focused on the impact of extracellular S100A11 in MPM and explored the therapeutic potential of an S100A11-targeting strategy. We examined the secretion level of S100A11 in various kinds of cell lines by enzyme-linked immunosorbent assay. Among them, six out of seven MPM cell lines actively secreted S100A11, whereas normal mesothelial cell lines did not secrete it. To investigate the role of secreted S100A11 in MPM, we inhibited its function by neutralizing S100A11 with an anti-S100A11 antibody. Interestingly, the antibody significantly inhibited the proliferation of S100A11-secreting MPM cells in vitro and in vivo. Microarray analysis revealed that several pathways including genes involved in cell proliferation were negatively enriched in the antibody-treated cell lines. In addition, we examined the secretion level of S100A11 in various types of pleural effusions. We found that the secretion of S100A11 was significantly higher in MPM pleural effusions, compared to others, suggesting the possibility for the use of S100A11 as a biomarker. In conclusion, our results indicate that extracellular S100A11 plays important roles in MPM and may be a therapeutic target in S100A11-secreting MPM.

    DOI: 10.1038/s41389-017-0017-3

    Web of Science

    Scopus

    PubMed

    researchmap

  • Crumbs3 promotes metastasis of colon cancer by regulating focal adhesion components Reviewed

    Iioka Hidekazu, Saito Ken, Sakaguchi Masakiyo, Morii Eiichi, Kondo Eisaku

    CANCER SCIENCE   109   1039   2018.1

  • Development of novel biologics for cancer metastasis via prevention of extracellular S100A8/A9 function Reviewed

    Kinoshita Rie, Yamauchi Akira, Shien Kazuhiko, Tomida Shuta, Toyooka Shinichi, Kondo Eisaku, Sakaguchi Masakiyo

    CANCER SCIENCE   109   1017   2018.1

  • Crucial role of RAGE in inappropriate increase of smooth muscle cells from patients with pulmonary arterial hypertension. Reviewed International journal

    Kazufumi Nakamura, Masakiyo Sakaguchi, Hiromi Matsubara, Satoshi Akagi, Toshihiro Sarashina, Kentaro Ejiri, Kaoru Akazawa, Megumi Kondo, Koji Nakagawa, Masashi Yoshida, Toru Miyoshi, Takeshi Ogo, Takahiro Oto, Shinichi Toyooka, Yuichiro Higashimoto, Kei Fukami, Hiroshi Ito

    PloS one   13 ( 9 )   e0203046   2018

     More details

    Language:English  

    BACKGROUND: Pulmonary vascular remodeling of pulmonary arterial hypertension (PAH) is characterized by an inappropriate increase of vascular cells. The receptor for advanced glycation end products (RAGE) is a type I single-pass transmembrane protein belonging to the immunoglobulin superfamily and is involved in a broad range of hyperproliferative diseases. RAGE is also implicated in the etiology of PAH and is overexpressed in pulmonary artery smooth muscle cells (PASMCs) in patients with PAH. We examined the role of RAGE in the inappropriate increase of PASMCs in patients with PAH. METHODS AND RESULTS: PASMCs were obtained from 12 patients with PAH including 9 patients with idiopathic PAH (IPAH) and 3 patients with heritable PAH (HPAH) (2 patients with BMPR2 mutation and one patient with SMAD9 mutation) who underwent lung transplantation. Western blot analysis and immunofluorescence staining revealed that RAGE and S100A8 and A9, ligands of RAGE, were overexpressed in IPAH and HPAH-PASMCs in the absence of any external growth stimulus. PDGF-BB (10 ng/mL) up-regulated the expression of RAGE in IPAH and HPAH-PASMCs. PAH-PASMCs are hyperplastic in the absence of any external growth stimulus as assessed by 3H-thymidine incorporation. This result indicates overgrowth characterized by continued growth under a condition of no growth stimulation in PAH-PASMCs. PDGF-BB stimulation caused a higher growth rate of PAH-PASMCs than that of non-PAH-PASMCs. AS-1, an inhibitor of TIR domain-mediated RAGE signaling, significantly inhibited overgrowth characterized by continued growth under a condition of no growth stimulation in IPAH and HPAH-PASMCs (P<0.0001). Furthermore, AS-1 significantly inhibited PDGF-stimulated proliferation of IPAH and HPAH-PASMCs (P<0.0001). CONCLUSIONS: RAGE plays a crucial role in the inappropriate increase of PAH-PASMCs. Inhibition of RAGE signaling may be a new therapeutic strategy for PAH.

    DOI: 10.1371/journal.pone.0203046

    PubMed

    researchmap

  • Stromal mesenchymal stem cells facilitate pancreatic cancer progression by regulating specific secretory molecules through mutual cellular interaction. Reviewed International journal

    Ken Saito, Masakiyo Sakaguchi, Satoshi Maruyama, Hidekazu Iioka, Endy Widya Putranto, I Wayan Sumardika, Nahoko Tomonobu, Takashi Kawasaki, Keiichi Homma, Eisaku Kondo

    Journal of Cancer   9 ( 16 )   2916 - 2929   2018

     More details

    Language:English  

    Pancreatic ductal adenocarcinoma (PDAC) is currently one of the most intractable malignancies with a typical scirrhous pattern in histology. Due to its abundant tumor stroma and scant vascularization, chemotherapeutic agents are considered inefficiently permeable to cancer nests, making it highly difficult to cure the patients with PDAC. However, PDAC is also considered to owe its intractability to other critical factors such as cellular interaction between tumor cells and tumor microenvironment as well as architectural barriers, which increases in therapeutic resistance. Here, we report a specific cellular interaction between PDAC cells and mesenchymal stem cells (MSCs) intermingled in PDAC stroma, which facilitates cancer invasion. Secretory phenotype profiling revealed that production of Amphiregulin (AREG) and MMP-3 were specifically upregulated under the coexistence of BxPC3 cells with human MSCs (approximately four to ten folds in AREG, and twenty to sixty-folds in MMP-3 compared to that of BxPC3 cells alone), whereas MMP-9 expression was decreased (less than one-tenth comparing with that of BxPC3 cells alone). Blockage of AREG production by its specific siRNA removed MSC-mediated driving force of BxPC3 invasiveness. Immunohistochemical analysis of tissue samples obtained both from PDAC patients and PDAC imitating mouse xenografted models revealed that significant coexpression of AREG and its receptor EGFR were detected on the cancer cells at invasive front. These results strongly suggested that cellular interaction between cancer cells and MSCs in the PDAC stroma might be critical to cancer progression, especially in the process of local invasion and the early stage development of metastasis.

    DOI: 10.7150/jca.24415

    Web of Science

    PubMed

    researchmap

  • Signal Diversity of Receptor for Advanced Glycation End Products Reviewed

    Masakiyo Sakaguchi, Rie Kinoshita, Endy Widya Putranto, I. Made Winarsa Ruma, I. Wayan Sumardika, Chen Youyi, Naoko Tomonobu, Ken-ichi Yamamoto, Hitoshi Murata

    ACTA MEDICA OKAYAMA   71 ( 6 )   459 - 465   2017.12

     More details

    Language:English   Publisher:OKAYAMA UNIV MED SCHOOL  

    The receptor for advanced glycation end products (RAGE) is involved in inflammatory pathogenesis. It functions as a receptor to multiple ligands such as AGEs, HMGB1 and S100 proteins, activating multiple intracellular signaling pathways with each ligand binding. The molecular events by which ligand-activated RAGE controls diverse signaling are not well understood, but some progress was made recently. Accumulating evidence revealed that RAGE has multiple binding partners within the cytoplasm and on the plasma membrane. It was first pointed out in 2008 that RAGE's cytoplasmic tail is able to recruit Diaphanous-1 (Dia-1), resulting in the acquisition of increased cellular motility through Rac1/Cdc42 activation. We also observed that within the cytosol, RAGE's cytoplasmic tail behaves similarly to a Toll-like receptor (TLR4)-TIR domain, interacting with TIRAP and MyD88 adaptor molecules that in turn activate multiple downstream signals. Subsequent studies demonstrated the presence of an alternative adaptor molecule, DAP10, on the plasma membrane. The coupling of RAGE with DAP10 is critical for enhancing the RAGE-mediated survival signal. Interestingly, RAGE interaction on the membrane was not restricted to DAP10 alone. The chemotactic G-protein-coupled receptors (GPCRs) formyl peptide receptors1 and 2 (FPR1 and FPR2) also interacted with RAGE on the plasma membrane. Binding interaction between leukotriene B4 receptor 1 (BLT1) and RAGE was also demonstrated. All of the interactions affected the RAGE signal polarity. These findings indicate that functional interactions between RAGE and various molecules within the cytoplasmic area or on the membrane area coordinately regulate multiple ligand-mediated RAGE responses, leading to typical cellular phenotypes in several pathological settings. Here we review RAGE's signaling diversity, to contribute to the understanding of the elaborate functions of RAGE in physiological and pathological contexts.

    DOI: 10.18926/AMO/55582

    Web of Science

    PubMed

    researchmap

  • Exogenous DKK-3/REIC inhibits Wnt/beta-catenin signaling and cell proliferation in human kidney cancer KPK1 Reviewed

    Jiaqi Xu, Takuya Sadahira, Rie Kinoshita, Shun-Ai Li, Peng Huang, Koichiro Wada, Motoo Araki, Kazuhiko Ochiai, Hirofumi Noguchi, Masakiyo Sakaguchi, Yasutomo Nasu, Masami Watanabe

    ONCOLOGY LETTERS   14 ( 5 )   5638 - 5642   2017.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    The third member of the Dickkopf family (DKK-3), also known as reduced expression in immortalized cells (REIC), is a tumor suppressor present in a variety of tumor cells. Regarding the regulation of the Wnt/beta-catenin signaling pathway, exogenous DKK-1 and DKK-2 are reported to inhibit Wnt signaling by binding the associated effectors. However, whether exogenous DKK-3 inhibits Wnt signaling remains unclear. A recombinant protein of human full-length DKK-3 was used to investigate the exogenous effects of the protein in vitro in KPK1 human renal cell carcinoma cells. It was demonstrated that the expression of phosphorylated (p-)beta-catenin (inactive form as the transcriptional factor) was increased in KPK1 cells treated with the exogenous DKK-3 protein. The levels of non-p-beta-catenin (activated form of beta- catenin) were consistently decreased. It was revealed that the expression of transcription factor (TCF) 1 and c-Myc, the downstream transcription factors of the Wnt/beta-catenin signaling pathway, was inhibited following treatment with DKK-3. A cancer cell viability assay confirmed the anti-proliferative effects of exogenous DKK-3 protein, which was consistent with a suppressed Wnt/beta-catenin signaling cascade. In addition, as low-density lipoprotein receptor-related protein 6 (LRP6) is a receptor of DKK-1 and DKK-2 and their interaction on the cell surface inhibits Wnt/beta-catenin signaling, it was examined whether the exogenous DKK-3 protein affects LRP6-mediated Wnt/beta-catenin signaling. The LRP6 gene was silenced and the effects of DKK-3 on the time course of the upregulation of p-beta-catenin expression were subsequently analyzed. Notably, LRP6 depletion elevated the base level of p-beta-catenin; however, there was no significant effect on its upregulation course or expression pattern. These findings indicate that exogenous DKK-3 upregulates p-beta-catenin and inhibits Wnt/beta-catenin signaling in an LRP6-independent manner. Therefore, exogenous DKK-3 protein may inhibit the proliferation of KPK1 cells via inactivating Wnt/beta-catenin signaling.

    DOI: 10.3892/ol.2017.6833

    Web of Science

    PubMed

    researchmap

  • Promising therapeutic efficacy of a novel reduced expression in immortalized cells/dickkopf-3 expressing adenoviral vector for hepatocellular carcinoma Reviewed

    Hiroaki Sawahara, Hidenori Shiraha, Daisuke Uchida, Hironari Kato, Ryo Kato, Atsushi Oyama, Teruya Nagahara, Masaya Iwamuro, Shigeru Horiguchi, Koichiro Tsutsumi, Mari Mandai, Tetsushige Mimura, Nozomu Wada, Yasuto Takeuchi, Kenji Kuwaki, Hideki Onishi, Shinichiro Nakamura, Masami Watanabe, Masakiyo Sakaguchi, Akinobu Takaki, Kazuhiro Nouso, Takahito Yagi, Yasutomo Nasu, Hiromi Kumon, Hiroyuki Okada

    JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY   32 ( 10 )   1769 - 1777   2017.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY  

    Background and Aim: Reduced expression in immortalized cells (REIC)/dickkopf-3 (Dkk-3) is a tumor suppressor gene that is downregulated in various cancers. In our previous study of prostate cancer, the REIC/Dkk-3-expressing adenoviral vector (Ad-REIC) was found to induce cancer-selective apoptosis. This study recently developed a novel super gene expression (SGE) system and used this system to re-construct an Ad-REIC vector, termed the Ad-SGE-REIC, to achieve more effective therapeutic outcomes. In this study, the therapeutic effects of Ad-SGE-REIC on hepatocellular carcinoma (HCC) was assessed.
    Methods: Human HCC cell lines (HLE, Huh7, HepG2, HLF, SK-Hep1, and PLC), human HCC tissues, and mouse HCC cell line (Hepa1-6) were used in this study. REIC/Dkk-3 expression was assessed by immunoblotting and immunohistochemistry. The relative cell viability and the apoptotic effect were examined in vitro, and the anti-tumor effects of Ad-SGE-REIC treatment were analyzed in the mouse xenograft model. This study additionally assessed anti-tumor immunological effects on the immunocompetent mice.
    Results: REIC/Dkk-3 expression was decreased in HCC cell lines and HCC tissues. Ad-SGE-REIC reduced cell viability and induced apoptosis in HCC cell lines (HLE and Huh7), inhibited tumor growth in the mouse xenograft model, and demonstrated in vivo anti-cancer immunostimulatory effects on the HCC cell line (Hepa1-6).
    Conclusions: Ad-SGE-REIC treatment not only enhanced cell killing effects in vitro but also elicited significant therapeutic effects, with tumor growth suppression, in vivo. REIC/Dkk-3 gene therapy using Ad-SGE-REIC potentially represents an innovative new therapeutic tool for HCC.

    DOI: 10.1111/jgh.13757

    Web of Science

    PubMed

    researchmap

  • Identification of a novel component leading to anti-tumor activity besides the major ingredient cordycepin in Cordyceps militaris extract Reviewed

    Takeharu Wada, I. Wayan Sumardika, Shingo Saito, I. Made Winarsa Ruma, Eisaku Kondo, Masami Shibukawa, Masakiyo Sakaguchi

    JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES   1061   209 - 219   2017.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE BV  

    In accordance with our previous study that was carried out to identify novel anti-tumor ingredients, chromatographic separation in combination with an anti-tumor activity assay was used for analysis of Cordyceps militaris extract in this study. Various modes of chromatography including reversed-phase, cation-exchange and anion exchange were used to separate components of Cordyceps militaris, which showed various chemical properties. Anti-tumor activity of each fraction was assessed by a Hoechst staining-based apoptosis assay using malignant melanoma MeWo cells. By these repeated approaches through chromatographic segregation and cell biological assay, we finally succeeded in identifying the target substance from a certain fraction that included neutral hydrophilic components using a pre-column and post-column chlorine adduct ionization LC APCI MS method. The target substance was a mono-carbohydrate, xylitol, that induced apoptotic cell death in MeWo cells but not in normal human OUMS-24 fibroblasts. This is the first study showing that Cordyceps militaris extract contains a large amount of xylitol. Thus, our results will contribute greatly to uncovering the mysterious multifunctional herbal drug Cordyceps militaris as an anti-tumor agent.

    DOI: 10.1016/j.jchromb.2017.07.022

    Web of Science

    PubMed

    researchmap

  • Robust cancer-specific gene expression by a novel cassette with hTERT and CMV promoter elements Reviewed

    Masakiyo Sakaguchi, Takuya Sadahira, Hideo Ueki, Rie Kinoshita, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Yasutomo Nasu, Kazuhiko Ochiai, Hiromi Kumon, Nam-Ho Huh, Masami Watanabe

    ONCOLOGY REPORTS   38 ( 2 )   1108 - 1114   2017.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    We developed and validated a novel hTERT/CMV promoter element-driven gene expression cassette that can robustly enhance cancer-specific gene expression. The following gene expressional elements were located in tandem within the plasmid construct: [hTERT core promoter, cytomegalovirus (CMV) minimized promoter, RU5' sequence, an inserted gene, BGH polyA, hTERT enhancer]; this is hereafter referred to as the hT/Cm-R-hT construct. Using various human cancer cell lines and normal cells, the cancer-specific transcription of the green fluorescent protein (GFP) gene was examined by western blotting and fluorescence microscopy. Cancer-specific gene expression was robustly achieved in the hT/Cm-R-hT plasmid in comparison to the other control hT/Cm-driven construct. Notably, the expression level of GFP observed in the hT/Cm-RhT-driven construct was superior to that of the control plasmid with the conventional CMV promoter in HEK293 cells, which are known to possess higher hTERT activity than normal cells. We next examined the availability of hT/Cm-R-hT in detecting the target GFP expressing cancer cells from human peripheral blood mononuclear cells (PBMCs). The hT/Cm-R-hT plasmid successfully induced cancer-specific gene expression; the robust expression of GFP was observed in target HeLa cancer cells, whereas GFP was not visibly expressed in normal PBMCs. The plasmid allowed for the selective visualization of viable HeLa cancer cells in mixed cell cultures containing up to 10000-fold more PBMCs. These findings indicate that the hT/Cm-R-hT expressional system is a valuable tool for detecting viable cancer cells mixed with normal cells. The current system can therefore be applied to the in vitro detection of cancer cells that are disseminated in the blood and other types of body fluid in vivo. Since the current system can also be applied to other types of vectors, including virus vectors, this approach using the hTERT promoter-based construct is expected to become a valuable tool for enhancing cancer specific gene expression.

    DOI: 10.3892/or.2017.5710

    Web of Science

    PubMed

    researchmap

  • Expression of tumor suppressor REIC/Dkk-3 by a newly improved adenovirus vector with insertion of a hTERT promoter at the 3'-side of the transgene Reviewed

    Endy Widya Putranto, Rie Kinoshita, Masami Watanabe, Takuya Sadahira, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Ken Kataoka, Yusuke Inoue, I. Made Winarsa Ruma, I. Wayan Sumardika, Chen Youyi, Miyoko Kubo, Yoshihiko Sakaguchi, Kenji Saito, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh, Masakiyo Sakaguchi

    ONCOLOGY LETTERS   14 ( 1 )   1041 - 1048   2017.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Reduced expression in immortalized cells (REIC)/Dickkopf-3 (Dkk-3) overexpression, induced using an adenovirus (Ad)-REIC, has been revealed to have a dramatic therapeutic effect on multiple types of cancer. To achieve an improved therapeutic effect from Ad-REIC on cancer, our group previously developed an enhanced gene expression system, the C-TSC cassette [cytomegalovirus (CMV)-RU5' located upstream (C); another promoter unit composed of triple tandem promoters, human telomerase reverse transcriptase (hTERT), simian virus 40 and CMV, located downstream of the cDNA (TSC); plus a polyadenylation (polyA) signal]. When applied to the conventional Ad-REIC, this novel system induced the development of an enhanced product, Ad-C-TSC-REIC, which exhibited a noticeable anticancer effect. However, there were difficulties in terms of Ad-C-TSC-REIC productivity in HEK293 cells, which are a widely used donor cell line for viral production. Productivity of Ad-C-TSC-REIC was significantly reduced compared with the conventional Ad-REIC, as the Ad-C-TSC-REIC had a significantly higher ability to induce apoptotic cell death of not only various types of cancer cell, but also HEK293 cells. The present study aimed to overcome this problem by modifying the C-TSC structure, resulting in an improved candidate: A C-T cassette (C: CMV-RU5' located upstream; T: another promoter unit composed of a single hTERT promoter, located downstream of the cDNA plus a polyA signal), which demonstrated gene expression comparable to that of the C-TSC system. The improved adenovirus REIC/Dkk-3 product with the C-T cassette, named Ad-C-T-REIC, exhibited a higher expression level of REIC/Dkk3, similar to that of Ad-C-TSC-REIC. Notably, the vector mitigated the cell death of donor HEK293 cells, resulting in a higher rate of production of its adenovirus. These results indicated that Ad-C-T-REIC has the potential to be a useful tool for application in cancer gene therapy.

    DOI: 10.3892/ol.2017.6201

    Web of Science

    PubMed

    researchmap

  • An HNF4-microRNA-194/192 signaling axis maintains hepatic cell function Reviewed

    Aoi Morimoto, Mana Kannari, Yuichi Tsuchida, Shota Sasaki, Chinatsu Saito, Tsuyoshi Matsuta, Tsukasa Maeda, Megumi Akiyama, Takahiro Nakamura, Masakiyo Sakaguchi, Nobukazu Nameki, Frank J. Gonzalez, Yusuke Inoue

    JOURNAL OF BIOLOGICAL CHEMISTRY   292 ( 25 )   10574 - 10585   2017.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Hepatocyte nuclear factor 4 (HNF4) controls the expression of liver-specific protein-coding genes. However, some microRNAs are also modulated by HNF4, and it is not known whether they are direct targets of HNF4 and whether they influence hepatic function. In this study, we found that HNF4 regulates microRNAs, indicated by marked down-regulation of miR-194 and miR-192 (miR-194/192) in liver-specific Hnf4a-null (Hnf4a(H)) mice. Transactivation of the shared miR-194/192 promoter was dependent on HNF4 expression, indicating that miR-194/192 is a target gene of HNF4. Screening of potential mRNAs targeted by miR-194/192 revealed that expression of genes involved in glucose metabolism (glycogenin 1 (Gyg1)), cell adhesion and migration (activated leukocyte cell adhesion molecule (Alcam)), tumorigenesis and tumor progression (Rap2b and epiregulin (Ereg)), protein SUMOylation (Sumo2), epigenetic regulation (Setd5 and Cullin 4B (Cln4b)), and the epithelial-mesenchymal transition (moesin (Msn)) was up-regulated in Hnf4a(H) mice. Moreover, we also found that miR-194/192 binds the 3-UTR of these mRNAs. siRNA knockdown of HNF4 suppressed miR-194/192 expression in human hepatocellular carcinoma (HCC) cells and resulted in up-regulation of their mRNA targets. Inhibition and overexpression experiments with miR-194/192 revealed that Gyg1, Setd5, Sumo2, Cln4b, and Rap2b are miR-194 targets, whereas Ereg, Alcam, and Msn are miR-192 targets. These findings reveal a novel HNF4 network controlled by miR-194/192 that may play a critical role in maintaining the hepatocyte-differentiated state by inhibiting expression of genes involved in dedifferentiation and tumorigenesis. These insights may contribute to the development of diagnostic markers for early HCC detection, and targeting of the miR-194/192 pathway could be useful for managing HCC.

    DOI: 10.1074/jbc.M117.785592

    Web of Science

    PubMed

    researchmap

  • Distant Bystander Effect of REIC/DKK3 Gene Therapy Through Immune System Stimulation in Thoracic Malignancies Reviewed

    Ken Suzawa, Kazuhiko Shien, Huang Peng, Masakiyo Sakaguchi, Masami Watanabe, Shinsuke Hashida, Yuho Maki, Hiromasa Yamamoto, Shuta Tomida, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Hiromi Kumon, Shinichiro Miyoshi, Shinichi Toyooka

    ANTICANCER RESEARCH   37 ( 1 )   301 - 307   2017.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:INT INST ANTICANCER RESEARCH  

    Background: Reduced expression in immortalized cell (REIC)/Dickkoph-3 (DKK3) is a tumor-suppressor gene, and its overexpression by adenovirus vector (Ad-REIC) exhibits a remarkable therapeutic effect on various human cancer types through a mechanism triggered by endoplasmic reticulum stress. Materials and Methods: We examined the direct anti-tumor effect of Ad-REIC gene therapy on lung cancer and malignant mesothelioma cell lines in vitro, and the distant bystander effect using immunocompetent mouse allograft models with bilateral flank tumors. Results: Ad-REIC treatment showed antitumor effect in many lung cancer and malignant mesothelioma cell lines in vitro. In an in vivo model, Ad-REIC treatment inhibited the growth not only of directly treated tumors but also of distant untreated tumors. By immunohistochemical analysis, infiltration of T-cells and natural killer (NK) cells and expression of the major histocompatibility complex (MHC) class I molecules were observed in bilateral tumors. Conclusion: Ad-REIC treatment not only had a direct antitumor effect but also an indirect bystander effect through stimulation of the immune system.

    DOI: 10.21873/anticanres.11321

    Web of Science

    PubMed

    researchmap

  • S100-SPECT uncovers cellular and molecular events of pre-metastatic niche formation and following organ-specific cancer metastasis Reviewed

    Masakiyo Sakaguchi

    THERANOSTICS   7 ( 10 )   2649 - 2651   2017

     More details

    Language:English   Publisher:IVYSPRING INT PUBL  

    Great progress has been made in in vivo imaging for cancer metastasis. Eisenblaetter et al. developed an innovative S100A8/A9-specific single photon emission computed tomography (SPECT) probe for imaging and succeeded in detecting the metastatic organ favored by the cancer before the cancer arrival. By utilizing this advanced method, researchers have also found that cancer-derived monocytes are the main source of the abundant production of S100A8/A9 in the pre-metastatic area. The CCL2-CCR2 axis is associated with S100A8/A9 production. Clinical establishment of this technique is expected to enable accurate prediction and monitoring of cancer metastasis during therapy.

    DOI: 10.7150/thno.19866

    Web of Science

    PubMed

    researchmap

  • The 2016 Incentive Award of the Okayama Medical Association in General Medical Science (2016 Yuuki Prize)

    Sakaguchi Masakiyo

    Okayama Igakkai Zasshi (Journal of Okayama Medical Association)   129 ( 2 )   81 - 83   2017

     More details

    Language:Japanese   Publisher:Okayama Medical Association  

    DOI: 10.4044/joma.129.81

    CiNii Article

    CiNii Books

    researchmap

  • Crucial Role of RAGE in Inappropriate Increase of Smooth Muscle Cells From Patients With Pulmonary Arterial Hypertension Reviewed

    Kazufumi Nakamura, Satoshi Akagi, Toshihiro Sarashina, Masakiyo Sakaguchi, Hiromi Matsubara, Hiroshi Ito

    CIRCULATION   134   2016.11

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Web of Science

    researchmap

  • PINK1 Regulation of Mitochondrial Homeostasis and Cell Survival. Reviewed

    Hitoshi Murata, Ken-ichi Yamamoto, Rie Kinoshita, Ken Kataoka, Nam-ho Huh, Masakiyo Sakaguchi

    IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL   52   S8 - S8   2016.6

     More details

    Language:English   Publisher:SPRINGER  

    Web of Science

    researchmap

  • Development of a Novel REIC/DKK-3-Encoding Adenoviral Agent: Its Robust and Promising Therapeutic Effects in Pancreatic Cancer Reviewed

    Daisuke Uchida, Hidenori Shiraha, Hironari Kato, Sawahara Hiroaki, Masakiyo Sakaguchi, Masami Watanabe, Yasutomo Nasu, Hiromi Kumon, Hiroyuki Okada

    GASTROENTEROLOGY   150 ( 4 )   S294 - S294   2016.4

     More details

    Language:English   Publisher:W B SAUNDERS CO-ELSEVIER INC  

    DOI: 10.1016/S0016-5085(16)31025-3

    Web of Science

    researchmap

  • Receptor for advanced glycation endproducts modulates leukotriene B-4 receptor 1 signaling Reviewed

    Takako Ichiki, Tomoaki Koga, Toshiaki Okuno, Kazuko Saeki, Yasuhiko Yamamoto, Masakiyo Sakaguchi, Takehiko Yokomizo

    FASEB JOURNAL   30   2016.4

     More details

    Language:English   Publisher:FEDERATION AMER SOC EXP BIOL  

    DOI: 10.1089/dna.2016.3552

    Web of Science

    researchmap

  • Promising Therapeutic Efficacy of a Novel REIC/Dkk-3 Expressing Adenoviral Vector for Hepatocellular Carcinoma Reviewed

    Sawahara Hiroaki, Hidenori Shiraha, Daisuke Uchida, Masakiyo Sakaguchi, Masami Watanabe, Yasutomo Nasu, Hiromi Kumon, Hiroyuki Okada

    GASTROENTEROLOGY   150 ( 4 )   S1153 - S1153   2016.4

     More details

    Language:English   Publisher:W B SAUNDERS CO-ELSEVIER INC  

    DOI: 10.1016/S0016-5085(16)33891-4

    Web of Science

    researchmap

  • Receptor for advanced glycation endproducts modulates leukotriene B-4 receptor 1 signaling Reviewed

    Takako Ichiki, Tomoaki Koga, Toshiaki Okuno, Kazuko Saeki, Yasuhiko Yamamoto, Masakiyo Sakaguchi, Takehiko Yokomizo

    FASEB JOURNAL   30   2016.4

     More details

    Language:English   Publisher:FEDERATION AMER SOC EXP BIOL  

    DOI: 10.1089/dna.2016.3552

    Web of Science

    researchmap

  • Hepatocyte Nuclear Factor 4 alpha Controls Iron Metabolism and Regulates Transferrin Receptor 2 in Mouse Liver Reviewed

    Shunsuke Matsuo, Masayuki Ogawa, Martina U. Muckenthaler, Yumiko Mizui, Shota Sasaki, Takafumi Fujimura, Masayuki Takizawa, Nagayuki Ariga, Hiroaki Ozaki, Masakiyo Sakaguchi, Frank J. Gonzalez, Yusuke Inoue

    JOURNAL OF BIOLOGICAL CHEMISTRY   290 ( 52 )   30855 - 30865   2015.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Iron is an essential element in biological systems, but excess iron promotes the formation of reactive oxygen species, resulting in cellular toxicity. Several iron-related genes are highly expressed in the liver, a tissue in which hepatocyte nuclear factor 4 alpha (HNF4 alpha) plays a critical role in controlling gene expression. Therefore, the role of hepatic HNF4 alpha in iron homeostasis was examined using liver-specific HNF4 alpha-null mice (Hnf4a(Delta H) mice). Hnf4a(Delta H) mice exhibit hypoferremia and a significant change in hepatic gene expression. Notably, the expression of transferrin receptor 2 (Tfr2) mRNA was markedly decreased in Hnf4a(Delta H) mice. Promoter analysis of the Tfr2 gene showed that the basal promoter was located at a GC-rich region upstream of the transcription start site, a region that can be transactivated in an HNF4 alpha-independent manner. HNF4 alpha-dependent expression of Tfr2 was mediated by a proximal promoter containing two HNF4 alpha-binding sites located between the transcription start site and the translation start site. Both the GC-rich region of the basal promoter and the HNF4 alpha-binding sites were required for maximal transactivation. Moreover, siRNA knockdown of HNF4 alpha suppressed TFR2 expression in human HCC cells. These results suggest that Tfr2 is a novel target gene for HNF4 alpha, and hepatic HNF4 alpha plays a critical role in iron homeostasis.

    DOI: 10.1074/jbc.M115.694414

    Web of Science

    researchmap

  • NRF2 Regulates PINK1 Expression under Oxidative Stress Conditions Reviewed

    Hitoshi Murata, Hitoshi Takamatsu, Sulai Lie, Ken Kataoka, Nam-ho Huh, Masakiyo Sakaguchi

    PLoS One   10 ( 11 )   e0142438   2015.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PUBLIC LIBRARY SCIENCE  

    Mutations of the PTEN-induced putative kinase 1 (PINK1) gene are a cause of autosomal recessive forms of Parkinson's disease. Recent studies have revealed that PINK1 is an essential factor for controlling mitochondrial quality, and that it protects cells from oxidative stresses. Although there has been considerable progress in the elucidation of various aspects of PINK1 protein regulation such as activation, stability and degradation, the transcriptional regulation of PINK1 mRNA under stress conditions remains unclear. In this study, we found that nuclear factor (erythroid-derived 2)-like 2 (NRF2), an antioxidant transcription factor, regulates PINK1 expression under oxidative stress conditions. Damaged mitochondria arising from stress conditions induced NRF2-dependent transcription of the PINK1 gene through production of reactive oxygen species (ROS). Either an ROS scavenger or forced expression of KEAP1, a potent inhibitory partner to NRF2, restricted PINK1 expression induced by activated NRF2. Transcriptionally up-regulated PINK1 diminished oxidative stress-associated cell death. The results indicate that PINK1 expression is positively regulated by NRF2 and that the NRF2-PINK1 signaling axis is deeply involved in cell survival.

    DOI: 10.1371/journal.pone.0142438

    Web of Science

    PubMed

    researchmap

  • REIC/Dkk-3遺伝子治療の胸部悪性腫瘍に対する免疫刺激を介する抗腫瘍効果

    諏澤 憲, 枝園 和彦, 阪口 政清, 渡部 昌実, 橋田 真輔, 宗 淳一, 山本 寛斉, 牧 祐歩, 佃 和憲, 那須 保友, 公文 裕巳, 三好 新一郎, 豊岡 伸一

    日本癌学会総会記事   74回   J - 1224   2015.10

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • T790M変異を含むEGFR遺伝子変異を有する非小細胞肺癌細胞株に対するTAE226の抗腫瘍効果

    山本 寛斉, 阪口 政清, 宗 淳一, 佃 和憲, 木浦 勝行, 猶本 良夫, 三好 新一郎, 豊岡 伸一

    日本癌学会総会記事   74回   P - 2193   2015.10

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • TAE226, a Bis-Anilino Pyrimidine Compound, Shows Anti-Tumor Effect on EGFR-Mutant Non-Small Cell Lung Cancer Cells including T790M Mutant Reviewed

    Hiromasa Yamamoto, Hiroki Otani, Munenori Takaoka, Masakiyo Sakaguchi, Junichi Soh, Masaru Jida, Tsuyoshi Ueno, Takafumi Kubo, Hiroaki Asano, Kazunori Tsukuda, Katsuyuki Kiura, Shinji Hatakeyama, Eiji Kawahara, Yoshio Naomoto, Shinichiro Miyoshi, Shinichi Toyooka

    JOURNAL OF THORACIC ONCOLOGY   10 ( 9 )   S311 - S312   2015.9

     More details

    Language:English   Publisher:ELSEVIER SCIENCE INC  

    DOI: 10.1371/journal.pone.0129838

    Web of Science

    researchmap

  • Distant Bystander Effect of REIC/Dkk-3 Gene Therapy through Immune System Stimulation in a Murine Model of Thoracic Malignancies Reviewed

    Ken Suzawa, Kazuhiko Shien, Peng Huang, Masakiyo Sakaguchi, Masami Watanabe, Shinsuke Hashida, Junichi Soh, Hiromasa Yamamoto, Yuho Maki, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Hiromi Kumon, Shinichiro Miyoshi, Shinichi Toyooka

    JOURNAL OF THORACIC ONCOLOGY   10 ( 9 )   S599 - S599   2015.9

     More details

    Language:English   Publisher:ELSEVIER SCIENCE INC  

    Web of Science

    researchmap

  • Anti-tumor effect of afatinib, an irreversible EGFR/HER2 dual inhibitor, in lung cancers harboring HER2 oncogene Reviewed

    Ken Suzawa, Shinichi Toyooka, Masakiyo Sakaguchi, Tomoaki Ohtsuka, Mototsugu Watanabe, Shinsuke Hashida, Yuho Maki, Hiromasa Yamamoto, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Shinichiro Miyoshi

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-2588

    Web of Science

    researchmap

  • Identification of a novel binding protein playing a critical role in HER2 activation in lung cancer cells Reviewed

    Tomoaki Ohtsuka, Masakiyo Sakaguchi, Katsuyoshi Takata, Shinsuke Hashida, Mototsugu Watanabe, Ken Suzawa, Yuho Maki, Hiromasa Yamamoto, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Shinichiro Miyoshi, Shinichi Toyooka

    CANCER RESEARCH   75   2015.8

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2015-48

    Web of Science

    researchmap

  • Novel germline G660D mutation in HER2 gene detected by whole-exome sequencing can predispose a patient to developing familial lung adenocarcinoma Reviewed

    Hiromasa Yamamoto, Koichiro Higasa, Masakiyo Sakaguchi, Kazuhiko Shien, Junichi Soh, Koichi Ichimura, Masashi Furukawa, Shinsuke Hashida, Kazunori Tsukuda, Nagio Takigawa, Keitaro Matsuo, Katsuyuki Kiura, Sinichiro Miyoshi, Fumihiko Matsuda, Shinichi Toyooka

    CANCER RESEARCH   74 ( 19 )   2014.10

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-LB-291

    Web of Science

    researchmap

  • 家族性・孤発性肺腺癌におけるHER2膜貫通領域の新規遺伝子変異(Novel functional mutations in the transmembrane domain of HER2 gene in familial and sporadic lung adenocarcinomas)

    山本 寛斉, 日笠 幸一郎, 阪口 政清, 枝園 和彦, 宗 淳一, 市村 浩一, 佃 和憲, 瀧川 奈義夫, 松尾 恵太郎, 木浦 勝行, 三好 新一郎, 松田 文彦, 豊岡 伸一

    日本癌学会総会記事   73回   E - 2012   2014.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • がんマーカー、サイトケラチン19(CK19)機能の新展開 HER2活性化におけるCK19の役割(Critical role of Cytokeratin-19 in an oncogenic activation of HER2)

    大塚 智昭, 阪口 政清, 渡邉 元嗣, 諏澤 憲, 橋田 真輔, 牧 佑歩, 山本 寛斉, 宗 淳一, 浅野 博昭, 佃 和憲, 三好 新一郎, 豊岡 伸一

    日本癌学会総会記事   73回   E - 2071   2014.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • 新規HER2膜貫通部領域遺伝子変異の機能解析(Novel HER2 mutations in transmembrane dmain result in constitutive autophosphorylation of HER2)

    諏澤 憲, 阪口 政清, 山本 寛斉, 宗 淳一, 牧 佑歩, 橋田 真輔, 大塚 智昭, 渡邉 元嗣, 浅野 博昭, 佃 和憲, 三好 新一郎, 豊岡 伸一

    日本癌学会総会記事   73回   P - 1328   2014.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Extract of Cordyceps militaris inhibits angiogenesis and suppresses tumor growth of human malignant melanoma cells Reviewed

    I. Made Winarsa Ruma, Endy Widya Putranto, Eisaku Kondo, Risayo Watanabe, Ken Saito, Yusuke Inoue, Ken-Ichi Yamamoto, Susumu Nakata, Masaji Kaihata, Hitoshi Murata, Masakiyo Sakaguchi

    INTERNATIONAL JOURNAL OF ONCOLOGY   45 ( 1 )   209 - 218   2014.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Angiogenesis is essential for tumor development and metastasis. Among several angiogenic factors, vascular endothelial growth factor receptor (VEGF) is important for tumor-derived angiogenesis and commonly overexpressed in solid tumors. Thus, many antitumor strategies targeting VEGF have been developed to inhibit cancer angiogenesis, offering insights into the successful treatment of solid cancers. However, there are a number of issues such as harmful effects on normal vascularity in clinical trials. Taking this into consideration, we employed Cordyceps militaris as an antitumor approach due to its biological safety in vivo. The herbal medicinal mushroom Cordyceps militaris has been reported to show potential anticancer properties including anti-angiogenic capacity; however, its concrete properties have yet to be fully demonstrated. In this study, we aimed to elucidate the biological role of Cordyceps militaris extract in tumor cells, especially in regulating angiogenesis and tumor growth of a human malignant melanoma cell line. We demonstrated that Cordyceps militaris extract remarkably suppressed tumor growth via induction of apoptotic cell death in culture that links to the abrogation of VEGF production in melanoma cells. This was followed by mitigation of Akt1 and GSK-3 beta activation, while p38 alpha phosphorylation levels were increased. Extract treatment in mouse model xenografted with human melanoma cells resulted in a dramatic antitumor effect with downregulation of VEGF expression. The results suggest that suppression of tumor growth by Cordyceps militaris extract is, at least, mediated by its anti-angiogenicity and apoptosis induction capacities. Cordyceps militaris extract may be a potent antitumor herbal drug for solid tumors.

    DOI: 10.3892/ijo.2014.2397

    Web of Science

    PubMed

    researchmap

  • Dramatic Increase in Expression of a Transgene by Insertion of Promoters Downstream of the Cargo Gene Reviewed

    Masakiyo Sakaguchi, Masami Watanabe, Rie Kinoshita, Haruki Kaku, Hideo Ueki, Junichiro Futami, Hitoshi Murata, Yusuke Inoue, Shun-Ai Li, Peng Huang, Endy Widya Putranto, I. Made Winarsa Ruma, Yasutomo Nasu, Hiromi Kumon, Nam-ho Huh

    MOLECULAR BIOTECHNOLOGY   56 ( 7 )   621 - 630   2014.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:HUMANA PRESS INC  

    For expression of genes in mammalian cells, various vectors have been developed using promoters including CMV, EF-1 alpha, and CAG promoters and have been widely used. However, such expression vectors sometimes fail to attain sufficient expression levels depending on the nature of cargo genes and/or on host cell types. In the present study, we aimed to develop a potent promoter system that enables high expression levels of cargo genes ubiquitously in many different cell types. We found that insertion of an additional promoter downstream of a cargo gene greatly enhanced the expression levels. Among the constructs we tested, C-TSC cassette (C: CMV-RU5' located upstream; TSC: another promoter unit composed of triple tandem promoters, hTERT, SV40, and CMV, located downstream of the cDNA plus a polyadenylation signal) had the most potent capability, showing far higher efficiency than that of potent conventional vector systems. The results indicate that the new expression system is useful for production of recombinant proteins in mammalian cells and for application as a gene therapeutic measure.

    DOI: 10.1007/s12033-014-9738-0

    Web of Science

    PubMed

    researchmap

  • S100A11 is required for efficient plasma membrane repair and survival of invasive cancer cells Reviewed

    Jyoti K. Jaiswal, Stine P. Lauritzen, Luana Scheffer, Masakiyo Sakaguchi, Jakob Bunkenborg, Sanford M. Simon, Tuula Kallunki, Marja Jaattela, Jesper Nylandsted

    NATURE COMMUNICATIONS   5   3795   2014.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:NATURE PUBLISHING GROUP  

    Cell migration and invasion require increased plasma membrane dynamics and ability to navigate through dense stroma, thereby exposing plasma membrane to tremendous physical stress. Yet, it is largely unknown how metastatic cancer cells acquire an ability to cope with such stress. Here we show that S100A11, a calcium-binding protein upregulated in a variety of metastatic cancers, is essential for efficient plasma membrane repair and survival of highly motile cancer cells. Plasma membrane injury-induced entry of calcium into the cell triggers recruitment of S100A11 and Annexin A2 to the site of injury. We show that S100A11 in a complex with Annexin A2 helps reseal the plasma membrane by facilitating polymerization of cortical F-actin and excision of the damaged part of the plasma membrane. These data reveal plasma membrane repair in general and S100A11 and Annexin A2 in particular as new targets for the therapy of metastatic cancers.

    DOI: 10.1038/ncomms4795

    Web of Science

    PubMed

    researchmap

  • A novel gene expression system strongly enhances the anticancer effects of a REIC/Dkk-3-encoding adenoviral vector Reviewed

    Masami Watanabe, Masakiyo Sakaguchi, Rie Kinoshita, Haruki Kaku, Yuichi Ariyoshi, Hideo Ueki, Ryuta Tanimoto, Shin Ebara, Kazuhiko Ochiai, Junichiro Futami, Shun-Ai Li, Peng Huang, Yasutomo Nasu, Nam-Ho Huh, Hiromi Kumon

    ONCOLOGY REPORTS   31 ( 3 )   1089 - 1095   2014.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Gene expression systems with various promoters, including the cytomegalovirus (CMV) promoter, have been developed to increase the gene expression in a variety of normal and cancer cells. In particular, in the clinical trials of cancer gene therapy, a more efficient and robust gene expression system is required to achieve sufficient therapeutic outcomes. By inserting the triple translational enhancer sequences of human telomerase reverse transcriptase (hTERT), Simian virus 40 (SV40) and CMV downstream of the sequence of the BGH polyA, we were able to develop a novel gene expression system that significantly enhances the expression of the genes of interest. We termed this novel gene expression cassette the super gene expression (SGE) system, and herein verify the utility of the SGE cassette for a replication-deficient adenoviral vector. We newly developed an adenoviral vector expressing the tumor suppressor, reduced expression in immortalized cells (REIC)/Dickkopf-3 (Dkk-3), based on the CMV promoter-driven SGE system (Ad-SGE-REIC) and compared the therapeutic utility of Ad-SGE-REIC with that of the conventional adenoviral vectors (Ad-CMV-REIC or Ad-CAG-REIC). The results demonstrated that the CMV promoter-SGE system allows for more potent gene expression, and that the Ad-SGE-REIC is superior to conventional adenoviral systems in terms of the REIC protein expression and therapeutic effects. Since the SGE cassette can be applied for the expression of various therapeutic genes using various vector systems, we believe that this novel system will become an innovative tool in the field of gene expression and gene therapy.

    DOI: 10.3892/or.2013.2958

    Web of Science

    PubMed

    researchmap

  • Anti-Cancer Effects of REIC/Dkk-3-encoding Adenoviral Vector for the Treatment of Non-small Cell Lung Cancer Reviewed

    Kazuhiko Shien, Norimitsu Tanaka, Masami Watanabe, Junichi Soh, Masakiyo Sakaguchi, Keitaro Matsuo, Hiromasa Yamamoto, Masashi Furukawa, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Nam-Ho Huh, Shinichiro Miyoshi, Hiromi Kumon, Shinichi Toyooka

    PLOS ONE   9 ( 2 )   e87900   2014.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PUBLIC LIBRARY SCIENCE  

    Objectives: REIC/Dkk-3 is down-regulated in a broad range of human cancer cells and is considered to function as a tumor suppressor. We previously reported that REIC/Dkk-3-expressing adenovirus vector (Ad-REIC) induced endoplasmic reticulum (ER) stress and cancer-specific apoptosis in human prostate cancer. In this study, we examined the therapeutic impact of Ad-REIC on non-small cell lung cancer (NSCLC).
    Materials and Methods: We examined the anti-tumor effect of Ad-REIC on 25 NSCLC cell lines in vitro and A549 cells in vivo. Two of these cell lines were artificially established as EGFR-tyrosine kinase inhibitor (TKI) resistant sublines.
    Results: Ad-REIC-treatment inhibited the cell viability by 40% or more in 13 (52%) of the 25 cell lines at multiplicity of infection (MOI) of 20 (20 MOI). These cell lines were regarded as being highly sensitive cells. The cell viability of a nonmalignant immortalized cell line, OUMS-24, was not inhibited at 200 MOI of Ad-REIC. The effects of Ad-REIC on EGFR-TKI resistant sublines were equivalent to those in the parental cell lines. Here, we demonstrated that Ad-REIC treatment activated c-Jun N-terminal kinase (JNK) in NSCLC cell lines, indicating the induction of ER stress with GRP78/BiP (GRP78) up-regulation and resulting in apoptosis. A single intratumoral injection of Ad-REIC significantly inhibited the tumorigenic growth of A549 cells in vivo. As predictive factors of sensitivity for Ad-REIC treatment in NSCLC, we examined the expression status of GRP78 and coxsackievirus and adenovirus receptor (CAR). We found that the combination of the GRP78 and CAR expressional statuses may be used as a predictive factor for Ad-REIC sensitivity in NSCLC cells.
    Conclusion: Ad-REIC induced JNK activation and subsequent apoptosis in NSCLC cells. Our study indicated that Ad-REIC has therapeutic potential against NSCLC and that the expression statuses of GRP78 and CAR may predict a potential therapeutic benefit of Ad-REIC.

    DOI: 10.1371/journal.pone.0087900

    Web of Science

    PubMed

    researchmap

  • Preclinical Evaluation of MicroRNA-34b/c Delivery for Malignant Pleural Mesothelioma Reviewed

    Tsuyoshi Ueno, Shinichi Toyooka, Takuya Fukazawa, Takafumi Kubo, Junichi Soh, Hiroaki Asano, Takayuki Muraoka, Norimitsu Tanaka, Yuho Maki, Kazuhiko Shien, Masashi Furukawa, Masakiyo Sakaguchi, Hiromasa Yamamoto, Kazunori Tsukuda, Shinichiro Miyoshi

    ACTA MEDICA OKAYAMA   68 ( 1 )   23 - 26   2014.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    The microRNA-34s (miR-34s) have p53 response elements in their 5'-flanking regions and demonstrate tumor-suppressive functions. In malignant pleural mesothelioma (MPM), we previously reported that expression of miR-34b and miR-34c (miR-34b/c) was frequently downregulated by methylation in MPM cell lines and primary tumors. The forced overexpression of miR-34b/c showed significant antitumor effects with the induction of apoptosis in MPM cells. In this study, we examined the in vivo antitumor effects of miR-34b/c using adenovirus vector on MPM. We subcutaneously transplanted NCI-H290, a human MPM cell line, into BALB/C mice and injected adenovirus vector expressing miR-34b/c, luciferase driven by the cytomegalovirus promoter (Ad-miR-34b/c or Ad-Luc), or PBS control into tumors over 5mm in diameter. A statistically significant growth inhibition of the tumor volume was observed in the Ad-miR-34b/c group from day 6 onward compared to the Ad-Luc group. The inhibition rate of Ad-miR-34b/c, compared to the tumor volume treated with Ad-Luc, was 58.6% on day 10 and 54.7% on day13. Our results indicate that adenovirus-mediated miR-34b/c gene therapy could be useful for the clinical treatment of MPM.

    DOI: 10.18926/AMO/52140

    Web of Science

    PubMed

    researchmap

  • Novel Germline Mutation in the Transmembrane Domain of HER2 in Familial Lung Adenocarcinomas Reviewed

    Hiromasa Yamamoto, Koichiro Higasa, Masakiyo Sakaguchi, Kazuhiko Shien, Junichi Soh, Koichi Ichimura, Masashi Furukawa, Shinsuke Hashida, Kazunori Tsukuda, Nagio Takigawa, Keitaro Matsuo, Katsuyuki Kiura, Shinichiro Miyoshi, Fumihiko Matsuda, Shinichi Toyooka

    JNCI-JOURNAL OF THE NATIONAL CANCER INSTITUTE   106 ( 1 )   djt338   2014.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OXFORD UNIV PRESS INC  

    We encountered a family of Japanese descent in which multiple members developed lung cancer. Using whole-exome sequencing, we identified a novel germline mutation in the transmembrane domain of the human epidermal growth factor receptor 2 (HER2) gene (G660D). A novel somatic mutation (V659E) was also detected in the transmembrane domain of HER2 in one of 253 sporadic lung adenocarcinomas. Because the transmembrane domain of HER2 is considered to be responsible for the dimerization and subsequent activation of the HER family and downstream signaling pathways, we performed functional analyses of these HER2 mutants. Mutant HER2 G660D and V659E proteins were more stable than wild-type protein. Both the G660D and V659E mutants activated Akt. In addition, they activated p38, which is thought to promote cell proliferation in lung adenocarcinoma. Our findings strongly suggest that mutations in the transmembrane domain of HER2 may be oncogenic, causing hereditary and sporadic lung adenocarcinomas.

    DOI: 10.1093/jnci/djt338

    Web of Science

    PubMed

    researchmap

  • Histidine-rich glycoprotein prevents septic lethality through neutrophil regulation Reviewed

    Nishibori M, Wake H, Mori S, Liu K, Morioka Y, Teshigawara K, Sakaguchi M, Kuroda K, Takahashi H, Ohtsuka A, Yoshino T, Morimatsu H

    Critical Care   1 - 53   2014

  • Draft genome sequence of Clostridium botulinum type B strain Osaka05, isolated from an infant patient with botulism in Japan Reviewed

    Yoshihiko Sakaguchi, Koji Hosomi, Jumpei Uchiyama, Yoshitoshi Ogura, Kaoru Umeda, Masakiyo Sakaguchi, Tomoko Kohda, Masafumi Mukamoto, Naoaki Misawa, Shigenobu Matsuzaki, Tetsuya Hayashi, Shunji Kozaki

    Genome Announcements   2 ( 1 )   2014

     More details

    Publishing type:Research paper (scientific journal)   Publisher:1  

    © 2014 Sakaguchi et al. Clostridium botulinum strain Osaka05, which has been isolated from an infant patient with botulism in Japan, is the first strain producing botulinum neurotoxin subtype B6. Here, we report the draft genome sequence of C. botulinum Osaka05.

    DOI: 10.1128/genomeA.e01010-13

    Scopus

    PubMed

    researchmap

  • Inhibition of RAGE signaling through the intracellular delivery of inhibitor peptides by PEI cationization Reviewed

    Endy Widya Putranto, Hitoshi Murata, Ken-Ichi Yamamoto, Ken Kataoka, Hidenori Yamada, Jun-Ichiro Futami, Masakiyo Sakaguchi, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   32 ( 4 )   938 - 944   2013.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    The receptor for advanced glycation end products (RAGE) is a multi-ligand cell surface receptor and a member of the immunoglobulin superfamily. RAGE is involved in a wide range of inflammatory, degenerative and hyper-proliferative disorders which span over different organs by engaging diverse ligands, including advanced glycation end products, S100 family proteins, high-mobility group protein B1 (HMGB1) and amyloid beta. We previously demonstrated that the cytoplasmic domain of RAGE is phosphorylated upon the binding of ligands, enabling the recruitment of two distinct pairs of adaptor proteins, Toll-interleukin 1 receptor domain-containing adaptor protein (TIRAP) and myeloid differentiation protein 88 (MyD88). This engagement allows the activation of downstream effector molecules, and thereby mediates a wide variety of cellular processes, such as inflammatory responses, apoptotic cell death, migration and cell growth. Therefore, inhibition of the binding of TIRAP to RAGE may abrogate intracellular signaling from ligand-activated RAGE. In the present study, we developed inhibitor peptides for RAGE signaling (RAGE-I) by mimicking the phosphorylatable cytosolic domain of RAGE. RAGE-I was efficiently delivered into the cells by polyethylenimine (PEI) cationization. We demonstrated that RAGE-I specifically bound to TIRAP and abrogated the activation of Cdc42 induced by ligand-activated RAGE. Furthermore, we were able to reduce neuronal cell death induced by an excess amount of S100B and to inhibit the migration and invasion of glioma cells in vitro. Our results indicate that RAGE-I provides a powerful tool for therapeutics to block RAGE-mediated multiple signaling.

    DOI: 10.3892/ijmm.2013.1467

    Web of Science

    PubMed

    researchmap

  • SARM1 and TRAF6 bind to and stabilize PINK1 on depolarized mitochondria Reviewed

    Hitoshi Murata, Masakiyo Sakaguchi, Ken Kataoka, Nam-ho Huh

    MOLECULAR BIOLOGY OF THE CELL   24 ( 18 )   2772 - 2784   2013.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER SOC CELL BIOLOGY  

    Mutations in PTEN-induced putative kinase 1 (PINK1) or parkin cause autosomal recessive forms of Parkinson's disease. Recent work suggests that loss of mitochondrial membrane potential stabilizes PINK1 and that accumulated PINK1 recruits parkin from the cytoplasm to mitochondria for elimination of depolarized mitochondria, which is known as mitophagy. In this study, we find that PINK1 forms a complex with sterile alpha and TIR motif containing 1 (SARM1) and tumor necrosis factor receptor-associated factor 6 (TRAF6), which is important for import of PINK1 in the outer membrane and stabilization of PINK1 on depolarized mitochondria. SARM1, which is known to be an adaptor protein for Toll-like receptor, binds to PINK1 and promotes TRAF6-mediated lysine 63 chain ubiquitination of PINK1 at lysine 433. Down-regulation of SARM1 and TRAF6 abrogates accumulation of PINK1, followed by recruitment of parkin to damaged mitochondria. Some pathogenic mutations of PINK1 reduce the complex formation and ubiquitination. These results indicate that association of PINK1 with SARM1 and TRAF6 is an important step for mitophagy.

    DOI: 10.1091/mbc.E13-01-0016

    Web of Science

    PubMed

    researchmap

  • DOCK7 is a critical regulator of the RAGE-Cdc42 signaling axis that induces formation of dendritic pseudopodia in human cancer cells Reviewed

    Ken-Ichi Yamamoto, Hitoshi Murata, Endy Widya Putranto, Ken Kataoka, Akira Motoyama, Toshihiko Hibino, Yusuke Inoue, Masakiyo Sakaguchi, Nam-Ho Huh

    ONCOLOGY REPORTS   29 ( 3 )   1073 - 1079   2013.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Cellular migration is a fundamental process linked to cancer metastasis. Growing evidence indicates that the receptor for advanced glycation end products (RAGE) plays a pivotal role in this process. With regard to downstream signal transducers of RAGE, diaphanous-1 and activated small guanine nucleotide triphosphatases, Rac1 and Cdc42, have been identified. To obtain precise insight into the direct downstream signaling mechanism of RAGE, we screened for proteins interacting with the cytoplasmic domain of RAGE employing an immunoprecipitation-liquid chromatography coupled with an electrospray tandem mass spectrometry system. In the present study, we found that the cytoplasmic domain of RAGE interacted with an atypical DOCK180-related guanine nucleotide exchange factor, dedicator of cytokinesis protein 7 (DOCK7). DOCK7 bound to the RAGE cytoplasmic domain and transduced a signal to Cdc42, resulting in the formation of abundant highly branched filopodia-like protrusions, dendritic pseudopodia. Blocking of the function of DOCK7 greatly abrogated the formation of dendritic pseudopodia and suppressed cellular migration. These results indicate that DOCK7 functions as an essential and downstream regulator of RAGE-mediated cellular migration through the formation of dendritic pseudopodia.

    DOI: 10.3892/or.2012.2191

    Web of Science

    PubMed

    researchmap

  • S100A9 is a novel ligand of EMMPRIN that promotes melanoma metastasis Reviewed

    Toshihiko Hibino, Masakiyo Sakaguchi, Shoko Miyamoto, Mami Yamamoto, Akira Motoyama, Junichi Hosoi, Tadashi Shimokata, Tomonobu Ito, Ryoji Tsuboi, Nam-Ho Huh

    Cancer Research   73 ( 1 )   172 - 183   2013.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:1  

    The calcium-binding proteins S100A8 and S100A9 can dimerize to form calprotectin, the release of which during tissue damage has been implicated in inflammation and metastasis. However, receptor(s) mediating the physiologic and pathophysiologic effects of this damage-associated "danger signal" are uncertain. In this study, searching for candidate calprotectin receptors by affinity isolation-mass spectrometry, we identified the cell surface glycoprotein EMMPRIN/BASIGIN (CD147/BSG). EMMPRIN specifically bound to S100A9 but not S100A8. Induction of cytokines and matrix metalloproteases (MMP) by S100A9 was markedly downregulated in melanoma cells by attenuation of EMMPRIN. We found that EMMPRIN signaling used the TNF receptor-associated factor TRAF2 distinct from the known S100-binding signaling pathway mediated by RAGE (AGER). S100A9 strongly promoted migration when EMMPRIN was highly expressed, independent of RAGE, whereas EMMPRIN blockade suppressed migration by S100A9. Immunohistologic analysis of melanomas revealed that EMMPRIN was expressed at both the invasive edge of lesions and the adjacent epidermis, where S100A9 was also strongly expressed. In epidermal-specific transgenic mice, tail vein-injected melanoma accumulated in skin expressing S100A9 but not S100A8. Together, our results establish EMMPRIN as a receptor for S100A9 and suggest the therapeutic use in targeting S100A9-EMMPRIN interactions. ©2012 AACR.

    DOI: 10.1158/0008-5472.CAN-11-3843

    Scopus

    PubMed

    researchmap

  • Preclinical biodistribution and safety study of reduced expression in immortalized cells/Dickkopf-3-encoding adenoviral vector for prostate cancer gene therapy Reviewed

    Morito Sugimoto, Masami Watanabe, Haruki Kaku, Shun-Ai Li, Hirofumi Noguchi, Hideo Ueki, Masakiyo Sakaguchi, Nam-Ho Huh, Yasutomo Nasu, Hiromi Kumon

    ONCOLOGY REPORTS   28 ( 5 )   1645 - 1652   2012.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    The biodistribution and safety of adenoviral vectors encoding the human REIC/Dkk-3 tumor suppressor gene (Ad-REIC) were examined in this preclinical study for in situ prostate cancer gene therapy. First, the in vitro apoptotic effects of Ad-REIC in normal and cancer cells derived from the prostate and liver were examined. Significant apoptotic effects were observed at 100 MOI (multiplicity of infection) in prostate cancer cells (LNCaP, PC3) and hepatoma cells (HEP3B and HEPG2); however, no effects were seen in normal cells. To analyze the safety of intraprostatic Ad-REIC administration, the biodistribution and histology after Ad-REIC injection were evaluated in various organs of normal male C57BL6 mice. In a supporting study, vector dissemination following intravenous injection of Ad-REIC into tail veins was determined. To evaluate whether Ad-REIC was present in the collected tissue specimens, human REIC gene detection was performed using DNA-PCR. Intraprostatic treatment administered at lower doses showed vector biodistribution into the colon, urinary bladder and prostate. At higher doses, vector dissemination was observed in tissues more distant from the prostate, including the lung, thymus, heart, liver and adrenal gland. After intravenous injection a: Ad-REIC, dissemination was observed in the liver and spleen. These results indicate that the biodistribution of Ad-REIC is determined by the dose and route of administration. Although acute inflammatory effects were observed in the prostate after intraprostatic administration at higher doses, no abnormal histological findings were noted in the other tissues, including those of intravenously treated mice. Regarding the safety of Ad-REIC administration, no deaths and no signs of toxicity or unusual behavior were observed in the mice in any treatment group. Based on these preclinical experiments, adenovirus-mediated in situ REIC/Dkk-3 gene therapy is considered to be safe for use as a treatment for human prostate cancer.

    DOI: 10.3892/or.2012.2001

    Web of Science

    PubMed

    researchmap

  • DIRECT THERAPEUTIC EFFECT OF REIC/DKK-3-ENCODING ADENOVIRAL VECTOR FOR NON-SMALL CELL LUNG CANCER Reviewed

    Yamamoto Hiromasa, Tanaka Norimitsu, Toyooka Shinichi, Watanabe Masami, Sakaguchi Masakiyo, Soh Junichi, Shien Kazuhiko, Furukawa Masashi, Asano Hiroaki, Tsukuda Kazunori, Nasu Yasutomo, Huh Nam-ho, Kumon Hiromi, Miyoshi Shinichirou

    JOURNAL OF THORACIC ONCOLOGY   7 ( 11 )   S463   2012.11

  • REIC/Dkk-3-encoding adenoviral vector as a potentially effective therapeutic agent for bladder cancer Reviewed

    Takeshi Hirata, Masami Watanabe, Haruki Kaku, Yasuyuki Kobayashi, Hiroshi Yamada, Masakiyo Sakaguchi, Kohji Takei, Nam-Ho Huh, Yasutomo Nasu, Hiromi Kumon

    INTERNATIONAL JOURNAL OF ONCOLOGY   41 ( 2 )   559 - 564   2012.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Bladder cancer is one of the most common urogenital malignancies. The intravesical instillation of anticancer agents is an attractive strategy to treat a superficial lesion or floating/disseminated cancer cells after transurethral operation. An adenovirus carrying REIC/Dkk-3, a tumor suppressor gene (Ad-REIC), exhibits cancer-specific apoptotic effects in various types of cancer cells. The aim of the present study was to examine the potential of Ad-REIC as a therapeutic agent for bladder cancer. KK47 and RT4 human bladder cancer cells were sensitive to the Ad-REIC treatment for apoptosis induction, but some human bladder cancer cell lines (T24, J82 and TccSup) were resistant. Significant cell growth inhibition was observed when these resistant cancer cell lines were treated with Ad-REIC in a condition of floating cells, which is clinically observed after transurethral operation and becomes a cause of intravesical cancer dissemination. The therapeutic potential of Ad-REIC for the treatment of multidrug-resistant bladder cancer was investigated. The adriamycin-resistant KK47 bladder cancer cells (KK47/ADM), which also present multidrug resistance, showed induction of significant apoptosis following Ad-REIC treatment. The Ad-REIC treatment induced downregulation of P-glycoprotein in KK47/ADM cells and restored the sensitivity to doxorubicin (adriamycin). Ad-REIC suppressed P-glycoprotein expression in a c-Jun-NH2-kinase (JNK)-dependent manner. Therefore, the current study indicated two therapeutic aspects of the Ad-REIC agent against human bladder cancer cells, as an apoptosis inducer/cell growth inhibitor and as a sensitizer of chemotherapeutic agents in multidrug-resistant cancer cells. The intravesical instillation of Ad-REIC could be an attractive therapeutic method in human bladder cancer where the treatment of superficial lesions and floating/disseminated or multidrug-resistant cancer cells is necessary.

    DOI: 10.3892/ijo.2012.1503

    Web of Science

    PubMed

    researchmap

  • S100A7 promotes the migration and invasion of osteosarcoma cells via the receptor for advanced glycation end products Reviewed

    Ken Kataoka, Tomoyuki Ono, Hitoshi Murata, Mika Morishita, Ken-Ichi Yamamoto, Masakiyo Sakaguchi, Nam-Ho Huh

    ONCOLOGY LETTERS   3 ( 5 )   1149 - 1153   2012.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    Osteosarcoma is the most common malignant tumor of bone in childhood and adolescence. Despite intensive research for new therapies, the outcome in patients with metastasis remains extremely poor. S100 proteins are involved in the proliferation, cell cycle progression and metastasis of numerous malignant tumors, including osteosarcoma. In the present study, we identified S100A7 as a candidate to promote the migration of osteosarcoma cells. S100A7 promoted the migration and invasion of osteosarcoma cells as assayed in vitro. An in vitro pull-down assay revealed the binding of the recombinant S100A7 protein with its putative receptor, the receptor for advanced glycation end products (RAGE). The downregulation of RAGE by a specific siRNA markedly suppressed the migration and invasion of osteosarcoma cells. Furthermore, the matrix metalloproteinase activity of osteosarcoma cells was enhanced by S100A7 and suppressed by the downregulation of RAGE. These results indicate that S100A7 promotes the migration and invasion of osteosarcoma cells through RAGE. The S100A7-RAGE axis may thus be a new target for preventing the invasion and/or metastasis of osteosarcoma.

    DOI: 10.3892/ol.2012.612

    Web of Science

    PubMed

    researchmap

  • 非小細胞肺癌に対する新たな個別化治療戦略 REIC/Dkk-3遺伝子治療の可能性

    宗 淳一, 豊岡 伸一, 田中 則光, 渡部 昌実, 山本 寛斉, 阪口 政清, 許 南浩, 那須 保友, 公文 裕巳, 三好 新一郎

    日本呼吸器外科学会雑誌   26 ( 3 )   P17 - 01   2012.4

     More details

    Language:Japanese   Publisher:(NPO)日本呼吸器外科学会  

    researchmap

  • Partial sensitization of human bladder cancer cells to a gene-therapeutic adenovirus carrying REIC/Dkk-3 by downregulation of BRPK/PINK1 Reviewed

    Yu Jin, Hitoshi Murata, Masakiyo Sakaguchi, Ken Kataoka, Masami Watanabe, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    ONCOLOGY REPORTS   27 ( 3 )   695 - 699   2012.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    REIC/Dkk-3 is a tumor suppressor gene that was first identified as a gene downregulated in association with immortalization of normal human fibroblasts. We have demonstrated that an adenovirus carrying REIC/Dkk-3 (Ad-REIC) showed a tumor-specific killing effect on a wide range of cancers. However, some human cancers, bladder cancers in particular, are resistant to Ad-REIC. In this study, we investigated the combination effect of downregulation of BRPK/PINK1 (PINK1) and Ad-REIC on bladder cancer cells. Five bladder cancer cell lines among six cell lines examined were resistant to Ad-REIC. Among the cell lines, the resistance of two cell lines was probably due to low infection efficiency of the adenovirus. PINK1-specific siRNA remarkably downregulated Bcl-x(L) and TRAP1 proteins and upregulated BAX protein expression. Finally, downregulation of PINK1 partially sensitized the other three cell lines that were resistant to Ad-REIC. This sensitization was associated with increasing production of reactive oxygen species (ROS). These results indicate that PINK1 is one of the key molecules for the mitochondrial protection system and that PINK1 can be a new target molecule to sensitize bladder cancer cells that are resistant to Ad-REIC.

    DOI: 10.3892/or.2011.1543

    Web of Science

    PubMed

    researchmap

  • Preclinical Safety and Efficacy of in Situ REIC/Dkk-3 Gene Therapy for Prostate Cancer Reviewed

    Keiichiro Kawauchi, Masami Watanabe, Haruki Kaku, Peng Huang, Kasumi Sasaki, Masakiyo Sakaguchi, Kazuhiko Ochiai, Nam-ho Huh, Yasutomo Nasu, Hiromi Kumon

    ACTA MEDICA OKAYAMA   66 ( 1 )   7 - 16   2012.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    The preclinical safety and therapeutic efficacy of adenoviral vectors that express the REIC/Dkk-3 tumor suppressor gene (Ad-REIC) was examined for use in prostate cancer gene therapy. The Ad-human (h) and mouse (m) REIC were previously demonstrated to induce strong anti-cancer effects in vitro and in vivo, and we herein report the results of two in vivo studies. First, intra-tumor Ad-hREIC administration was examined for toxicity and therapeutic effects in a subcutaneous tumor model using the PC3 prostate cancer cell line. Second, intra-prostatic Ad-mREIC administration was tested for toxicity in normal mice. The whole-body and spleen weights, hematological and serum chemistry parameters, and histological evaluation of tissues from throughout the body were analyzed. Both experiments indicated that there was no significant difference in the examined parameters between the Ad-REIC-treated group and the control (PBS- or Ad-LacZ-treated) group. In the in vitro analysis using PC3 cells, a significant apoptotic effect was observed after Ad-hREIC treatment. Confirming this observation, the robust anti-tumor efficacy of Ad-hREIC was demonstrated in the in vivo subcutaneous prostate cancer model. Based on the results of these preclinical experiments, we consider the adenovirus-mediated REIC/Dkk-3 in situ gene therapy to be safe and useful for the clinical treatment of prostate cancer.

    DOI: 10.18926/AMO/48076

    Web of Science

    PubMed

    researchmap

  • A Novel Tumor Suppressor, REIC/Dkk-3 Gene Identified by Our In Vitro Transformation Model of Normal Human Fibroblasts Works as a Potent Therapeutic Anti-tumor Agent Reviewed

    Masakiyo Sakaguchi, Nam-ho Huh, Masayoshi Namba

    HUMAN CELL TRANSFORMATION: ROLE OF STEM CELLS AND THE MICROENVIRONMENT   720   209 - 215   2012

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPRINGER-VERLAG BERLIN  

    Reduced Expression in Immortalized Cell (REIC) was cloned by subtractive hybridization method as a gene whose expression is reduced in many human immortalized and neoplastic tumor cells. The REIC, when overexpressed by an adenovirus (Ad-REIC), exhibited a dramatic therapeutic effect on a wide variety of human cancers through a mechanism triggered by ER-stress-mediated JNK activation. In addition to this direct effect on cancer cells, Ad-REIC exerted another cytotoxicity on human cancers, an indirect host-mediated effect due to overproduction of IL-7 by mis-targeted normal cells. This "one-bullet two-arms" finding may lead to a powerful new therapeutic approach to the treatment of human cancers.

    DOI: 10.1007/978-1-4614-0254-1_17

    Web of Science

    PubMed

    researchmap

  • S100A11, a dual growth regulator of epidermal keratinocytes Reviewed

    Masakiyo Sakaguchi, Nam-ho Huh

    AMINO ACIDS   41 ( 4 )   797 - 807   2011.10

     More details

    Language:English   Publisher:SPRINGER  

    S100A11, a member of the family of S100 proteins, is a dimmer, each monomer of which has two EF-hands. Expression of S100A11 is ubiquitous in various tissues at different levels, with a high expression level in the skin. We have analyzed functions of S100A11 mainly in normal human keratinocytes (NHK) as a model cell system of human epithelial cells. High Ca(2+) and transforming growth factor-beta (TGF-beta), two representative growth suppressors for NHK, need a common S100A11-mediated pathway in addition to unique pathways (NFAT1-mediated pathway for high Ca(2+) and Smad-mediated pathway for TGF-beta) for exhibiting a growth inhibitory effect. S100A11 has another action point for growth suppression in NHK. Annexin A1 (ANXA1) complexed with S100A11 efficiently binds to and inhibits cytosolic phospholipase A2 (cPLA2), the activity of which is needed for the growth of NHK. On exposure of NHK to epidermal growth factor (EGF), ANXA1 is cleaved at 12Trp, and this truncated ANXA1 loses binding capacity to S100A11, resulting in maintenance of an active state of cPLA2. On the other hand, we found that S100A11 is actively secreted by NHK. Extracellular S100A11 acts on NHK to enhance the production of EGF family proteins, resulting in growth stimulation. These findings indicate that S100A11 plays a dual role in growth regulation, being suppressive in cells and being promotive from outside of cells.

    DOI: 10.1007/s00726-010-0747-4

    Web of Science

    PubMed

    researchmap

  • 悪性胸膜中皮腫に対するマイクロRNA34b/cを用いたアデノウイルス遺伝子治療の可能性(Anti-proliferative effect of microRNA-34b/c adenoviral gene transfer on malignant pleural mesotheliomas)

    宗 淳一, 豊岡 伸一, 久保 孝文, 上野 剛, 深澤 拓也, 阪口 政清, 佃 和憲, 浅野 博昭, 山本 寛斉, 許 南浩, 三好 新一郎

    日本癌学会総会記事   70回   267 - 267   2011.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Tumor suppressor REIC/Dkk-3 interacts with the dynein light chain, Tctex-1 Reviewed

    Kazuhiko Ochiai, Masami Watanabe, Hideo Ueki, Peng Huang, Yasuyuki Fujii, Yasutomo Nasu, Hirofumi Noguchi, Takeshi Hirata, Masakiyo Sakaguchi, Nam-ho Huh, Yuji Kashiwakura, Haruki Kaku, Hiromi Kumon

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   412 ( 2 )   391 - 395   2011.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    REIC/Dkk-3 is a member of the Dickkopf family proteins known as Wnt-antagonists, and REIC/Dkk-3 expression is downregulated in a broad range of cancer types. REIC/Dkk-3 acts as a tumor suppressor in multiple cancer cell lines by inducing apoptosis through endoplasmic reticulum (ER) stress signaling. However, the intracellular interaction partners of REIC/Dkk-3 have not been fully elucidated. By employing yeast two-hybrid screening, we identified the human dynein light chain, Tctex-1, as a novel interaction partner of REIC/Dkk-3. We further disclosed that the interaction involves the 136-157 amino acid region of REIC/Dkk-3 by using the mammalian two-hybrid system. Interestingly, this binding region of REIC/Dkk-3 with Tctex-1 contains an amino acid sequence motif [-(E) under bar -X-(G) under bar-(R) under bar-(R) under bar -X-(H) under bar-] which was previously reported as the Tctex-1 binding domain of dynein intermediate chain (DIC). Immunocytochemistry demonstrated that both REIC/Dkk-3 and Tctex-1 were localized around the ER of human fibroblasts, and the similar distribution pattern of the proteins suggests that their interaction occurs around the ER. This is the first study showing the interaction of a Dickkopf family protein with a dynein motor complex protein. The link between REIC/Dkk-3 and Tctex-1 may be of significance for understanding the molecular functions of the proteins in ER stress signaling and intracellular dynein motor dynamics, respectively. (C) 2011 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2011.07.109

    Web of Science

    PubMed

    researchmap

  • The impact of adenoviral gene transfer of microRNA-34b/c on malignant pleural mesotheliomas Reviewed

    Junichi Soh, Shinichi Toyooka, Takafumi Kubo, Takuya Fukazawa, Masakiyo Sakaguchi, Tsuyoshi Ueno, Kazunori Tsukuda, Hiroaki Asano, Hiromasa Yamamoto, Nam-ho Huh, Shinichiro Miyoshi

    CANCER RESEARCH   71   2011.4

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2011-4944

    Web of Science

    researchmap

  • マイクロRNA34b/cのメチル化による発現低下は悪性胸膜中皮腫の重要な分子病態である(Epigenetic silencing of microRNA-34b/c plays a pivotal role in pathogenesis of malignant mesothelioma pathogenesis)

    久保 孝文, 豊岡 伸一, 佃 和憲, 阪口 政清, 深澤 拓也, 宗 淳一, 浅野 博昭, 那須 保友, 岸本 卓巳, 松井 秀樹, 許 南浩, 三好 新一郎

    日本癌学会総会記事   69回   44 - 44   2010.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Frequent silencing of tumor suppressive MicroRNA-34b/c by aberrant methylation in malignant mesothelioma Reviewed

    Takafumi Kubo, Shinichi Toyooka, Kazunori Tsukuda, Hiroaki Asano, Masakiyo Sakaguchi, Junichi Soh, Tsuyoshi Ueno, Hiromasa Yamamoto, Masaomi Yamane, Takahiro Oto, Yasutomo Nasu, Hideki Matsui, Nam Ho Huh, Shinichro Miyoshi

    CANCER RESEARCH   70   2010.4

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM10-2081

    Web of Science

    researchmap

  • REIC/Dkk-3 stable transfection reduces the malignant phenotype of mouse prostate cancer RM9 cells Reviewed

    Jie Chen, Masami Watanabe, Peng Huang, Masakiyo Sakaguchi, Kazuhiko Ochiai, Yasutomo Nasu, Mamoru Ouchida, Nam-Ho Huh, Kenji Shimizu, Yuji Kashiwakura, Haruki Kaku, Hiromi Kumon

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   24 ( 6 )   789 - 794   2009.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SPANDIDOS PUBL LTD  

    The reduced expression in immortalized cells (REIC)/Dickkopf (Dkk)-3. a member of the Dkk gene family, is a tumor suppressor in a broad range of cancers. REIC/Dkk-3 transfected stable clones of mouse prostate cancer RM9 cells (RM9-REIC) and the empty vector-transfected control clone cells (RM9-EV) were established. Clones were used to evaluate the anti-cancer effects and a proteomics analysis of REIC/Dkk-3 continuous expression was performed. The RM9-REIC cells show a feeble appearance and the cell membrane shows irregular buds known as blebs. In vitro cell proliferation was significantly suppressed in RM9-REIC clones in comparison to the control. The apoptosis assay was done under standard culture conditions and RM9-REIC showed a higher incidence of apoptosis. The RM9-EV and RM9-REIC cells were orthotopically implanted into a C57BL/6 mouse prostate. After 2 weeks, the tumor growth was significantly inhibited in RM9-REIC cells in comparison to the control. Two-dimensional gel electrophoresis was used to examine the modification of protein expression by the gene transfection. The analysis with mass spectrometry disclosed that expression of peroxiredoxin-l, GST-P1. transgelin-2, MRP-L12, ARD, GRP78 and Sorcin were increased and eEF1A-1 and cyclophilin-40 protein were decreased in RM9-REIC cells. Therefore, REIC/Dkk-3 stable transfectants show a reduction of malignancy in mouse prostate cancer RM9 cells in vitro and in vivo. The result of the proteomics analysis might provide important clues to clarify the anti-cancer molecular mechanism of REIC/Dkk-3 gene transfer.

    DOI: 10.3892/ijmm_00000293

    Web of Science

    PubMed

    researchmap

  • S100A11, a dual mediator for growth regulation of normal human keratinocytes Reviewed

    Masakiyo Sakaguchi, Nam-ho Huh

    AMINO ACIDS   37 ( 1 )   77 - 77   2009.7

     More details

    Language:English   Publisher:SPRINGER WIEN  

    Web of Science

    researchmap

  • Dual tyrosine kinase inhibitor for focal adhesion kinase and insulin-like growth factor-I receptor (TAE226) leads to apoptosis in esophageal cancer by inhibiting AKT-mTOR survival signaling Reviewed

    Huifang Hao, Zhigang Wang, Xiaohong Bao, Nobuyuki Watanabe, Kazufumi Sakurama, Yasuko Tomono, Takuya Fukazawa, Shinji Hatakeyama, Osamu Omori, Seishi Nishitani, Toshiaki Ohara, Munenori Takaoka, Yasuhiro Shirakawa, Tomoki Yamatsuji, Junji Matsuoka, Masakiyo Sakaguchi, Noriaki Tanaka, Yoshio Naomoto

    CANCER RESEARCH   69   2009.5

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    Web of Science

    researchmap

  • REIC/DKK-3 as a potential gene therapeutic agent against human testicular cancer Reviewed

    Ryuta Tanimoto, Fernando Abarzua, Masakiyo Sakaguchi, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    JOURNAL OF GENE MEDICINE   10 ( 4 )   454 - 454   2008.4

     More details

    Language:English   Publisher:JOHN WILEY & SONS LTD  

    Web of Science

    researchmap

  • Adenovirus-mediated REIC/DKK-3 gene transfer prevented mesothelioma tumor progressions in orthotopic mice model Reviewed

    Yuji Kashiwakura, Masami Watanabe, Fernando Abarzua, Masakiyo Sakaguchi, Munenori Takaoka, Ryuta Tanimoto, Yasutomo Nasu, Nam-ho Huh, Hiromi Kumon

    JOURNAL OF GENE MEDICINE   10 ( 4 )   469 - 470   2008.4

     More details

    Language:English   Publisher:JOHN WILEY & SONS LTD  

    Web of Science

    researchmap

  • Adenovirus-mediated overexpression of REIC/Dkk-3 selectively induces apoptosis in human prostate cancer cells through activation of c-Jun-NH2-kinase Reviewed

    F Abarzua, M Sakaguchi, M Takaishi, Y Nasu, K Kurose, S Ebara, M Miyazaki, M Namba, H Kumon, N Huh

    CANCER RESEARCH   65 ( 21 )   9617 - 9622   2005.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER ASSOC CANCER RESEARCH  

    Alteration in genes which takes place during malignant conversion and progression could be potential targets for gene therapy. We previously identified REIC/Dkk-3 as a gene whose expression is reduced in many human cancers. Here, we showed that expression of REIC/Dkk-3 was consistently reduced in human prostate cancer tissues in a stage-dependent manner. Forced expression of REIC/Dkk-3 induced apoptosis in human prostate cancer cell lines lacking endogenous REIC/Dkk-3 expression but not in REIC/Dkk3-proficient normal prostate epithelial and stromal cells. The apoptosis involved c-jun-NH2-kinase activation, mitochondrial translocation of Bax, and reduction of Bcl-2. A single injection of an adenovirus vector carrying REIC/Dkk-3 showed a dramatic antitumor effect on a xenotransplanted human prostate cancer. Thus, REIC/Dkk-3 could be a novel target for gene-based therapy of prostate cancer.

    DOI: 10.1158/0008-5472.CAN-05-0829

    Web of Science

    PubMed

    researchmap

  • NMR studies of the N-terminal segment of S100C/A11 protein Reviewed

    Takahide Kouno, Mineyuki Mizuguchi, Masakiyo Sakaguchi, Eiichi Makino, Nam-ho Huh, Keiichi Kawano

    Peptides 2004, Proceedings   1025 - 1026   2005

     More details

    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:KENES INT  

    Web of Science

    researchmap

  • Expression of CYP3A4 by an immortalized human hepatocyte line in a three-dimensional culture using a radial-flow bioreactor Reviewed

    Akiyama, I, K Tomiyama, M Sakaguchi, M Takaishi, M Mori, M Hosokawa, S Nagamori, N Shimizu, NH Huh, M Miyazaki

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   14 ( 4 )   663 - 668   2004.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PROFESSOR D A SPANDIDOS  

    Cytochrome P450 (CYP) 3A is responsible for about 50% of drug metabolizing activity in the liver. The present study was undertaken to establish a CYP3A4-active model for in vitro analysis of human drug metabolism. The cells used were immortalized normal human fetal hepatocytes (OUMS-29) and its HNF4alpha-introduced subline (OUMS-29/H-11). The cells were cultivated under high-density three-dimensional conditions in a radial-flow bioreactor (RFB). The number of OUMS-29 cells increased 15-fold over 49 days and their apical surfaces were covered with abundant microvilli, a characteristic of hepatocytes in vivo. The amount of albumin secreted by OUMS-29 cells in the three-dimensional RFB culture was 6-fold higher than those in a monolayer culture. CYP3A4 protein and an intermediate metabolite of testosterone by CYP3A4 were detected in OUMS-29/H11 cells cultivated in RFB &gt;29 days. These results indicate that the RFB; culture of OUMS-29/H-11 cells is useful for screening and developing new drugs.

    Web of Science

    PubMed

    researchmap

  • Involvement of interferon regulatory factor 1 and S100C/A11 in growth inhibition by transforming growth factor beta 1 in human hepatocellular carcinoma cells Reviewed

    M Miyazaki, M Sakaguchi, Akiyama, I, Y Sakaguchi, S Nagamori, NH Huh

    CANCER RESEARCH   64 ( 12 )   4155 - 4161   2004.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER ASSOC CANCER RESEARCH  

    Growth inhibition by transforming growth factor (TGF)-beta1 has been attributed to the induction of cyclin-dependent kinase inhibitors, among which p21/Waf1 plays a major role in many biological contexts. In the present study, two new intracellular mediators for the induction of p21/Waf1 by TGF-beta1 were identified in a human hepatocellular carcinoma cell line (JHH-5) expressing mutant-type p53. After addition of TGF-beta1 to JHH-5 cells, a marked increase of the p21/Waf1 expression preceded the inhibition of DNA synthesis. Expression of IFN regulatory factor (IRF)-1, a known transacting factor for p21/Waf1 promoter, was elevated just before or in parallel with the increase of p21/Waf1. Transduction of antisense IRF-1 inhibited the increase in p21/Waf1 in JHH-5 cells treated with TGF-beta1 and partially released the cells from the growth arrest by TGF-beta1. Expression of S100C/A11, a member of the Ca2+-binding S100 protein family, also markedly increased after addition of TGF-beta1. S100C/A11 protein was translocated to and accumulated in nuclei of TGF-beta1-treated JHH-5 cells, where p21/Waf1 was concomitantly accumulated. When a recombinant S100C/A11 protein was introduced into nuclei of JHH-5 cells, DNA synthesis was markedly inhibited in a dose-dependent manner in the absence of TGF-beta1. Prior transfection of p21/Waf1-targeted small interfering RNA efficiently blocked decrease of DNA synthesis in JHH-5 cells caused by TAT-S100C/A11 or TGF-beta1 and markedly inhibited expression of p21/Waf1 protein in the cells. These results indicate that IRF-1 and S100C/A11 mediate growth inhibition by TGF-beta1 via induction of p21/Waf1.

    DOI: 10.1158/0008-5472.CAN-03-2750

    Web of Science

    PubMed

    researchmap

  • Differential expression of S100C in thyroid lesions Reviewed

    C Torres-Cabala, A Panizo-Santos, HC Krutzsch, H Barazi, M Namba, M Sakaguchi, DD Roberts, MJ Merino

    INTERNATIONAL JOURNAL OF SURGICAL PATHOLOGY   12 ( 2 )   107 - 115   2004.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WESTMINSTER PUBL INC  

    Identification of new potential markers that may help in the diagnosis of benign and malignant thyroid lesions is needed. By comparative 2-dimensional gel electrophoresis of microdissected cells from tumors and normal thyroid tissue, we identified a new protein, S100C, which is highly expressed in papillary carcinomas. In order to validate this finding, we investigated the immunohistochemical expression and the potential role in diagnosis of these markets in 94 specimens representing the spectrum of malignant and benign thyroid lesions. Normal thyroid tissue was evaluated in 57 specimens. Galectin-3, a marker reported as specific for malignant lesions, was also evaluated in the same lesions. S100C protein was expressed in the nuclei of normal tissue, hyperplastic nodules, and follicular adenomas and carcinomas. Papillary carcinomas showed a strong, but cytoplasmic, pattern of staining. Galectin-3 immunostaining was strongly positive in papillary carcinomas, and negative in benign lesions, confirming its value in differential diagnosis. These findings Suggest that immunohistochemical Staining of S100C could be helpful in the pathological study of thyroid lesions, especially in cases in which follicular variants of papillary carcinoma and follicular carcinoma are considered in the differential diagnosis.

    DOI: 10.1177/106689690401200203

    Web of Science

    PubMed

    researchmap

  • PKC alpha mediates TGF beta-induced growth inhibition of human keratinocytes via phosphorylation of S100C/A11 Reviewed

    M Sakaguchi, M Miyazaki, H Sonegawa, M Kashiwagi, M Ohba, T Kuroki, M Namba, N Huh

    JOURNAL OF CELL BIOLOGY   164 ( 7 )   979 - 984   2004.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ROCKEFELLER UNIV PRESS  

    Growth regulation of epithelial cells is of major concern because most human cancers arise from them. We demonstrated previously a novel signal pathway involving S100C/A11 for high Ca2+-induced growth inhibition of normal human keratinocytes (Sakaguchi, M., M. Miyazaki, M. Takaishi, Y. Sakaguchi, E. Makino, N. Kataoka, H. Yamada, M. Namba, and N.H. Huh. 2003.J. Cell Biol. 163:825-835). This paper addresses a question whether transforming growth factor beta (TGFbeta) shares the pathway with high Ca2+. On exposure of the cells to TGFbeta1, S100C/A11 was phosphorylated, bound to nucleolin, and transferred to the nucleus, resulting in induction of p21(WAF1/CIP1) and p15(INK4B) through activation of Sp1. Protein kinase C alpha (PKCalpha) was shown to phosphorylate. (10)Thr of S100C/A11, which is a critical event for the signal transduction. The TGFbeta1-induced growth inhibition was almost completely mitigated when PKCalpha activity was blocked or when S100C/A11 was functionally sequestered. These results indicate that, in addition to the well-characterized Smad-mediated pathway, the PKCalpha-S100C/A11-mediated pathway is involved in and essential for the growth inhibition of normal human keratinocytes cells by TGFbeta1.

    DOI: 10.1083/jcb.200312041

    Web of Science

    PubMed

    researchmap

  • Lentiviral vector: A useful tool for transduction of human liver endothelial cells Reviewed

    T Totsugawa, N Kobayashi, M Maruyama, Y Kosaka, T Okitsu, T Arata, M Sakaguchi, T Ueda, Y Kurabayashi, N Tanaka

    ASAIO JOURNAL   49 ( 6 )   635 - 640   2003.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Endothelial cells play multiple roles in pathophysiologic processes and are increasingly being recognized as target cells of gene therapy. Lentiviral vectors derived from human immunodeficiency virus type 1 have an ability to infect both dividing and nondividing cells and currently receive a great deal of attention as an innovative tool for transduction of target cells. The purpose of the present work was to evaluate the efficacy of a lentiviral vector for transducing human liver endothelial cells (HLECs) in vitro. For the present study, a pseudotyped lentiviral vector encoding a green fluorescent protein (GFP) gene, LtV-GFP, was generated by means of FuGENE 6 method and allowed to infect HLECs. Approximately 95% of HLECs were positive for GFP expression after LtV-GFP infection at a multiplicity of infection of 10. Notably, LtV-GFP transduced HLECs had stable and long term GFP expression, showed gene expression of endothelial markers including CD 34, factor VIII, flt-1, KDR/flk-1 and HGF, and maintained in vitro angiogenic potential in a Matrigel assay to the same extent as primarily cultured HLECs. These findings provide evidence that lentivirus based gene delivery is an efficient tool for transduction of endothelial cells that could be considered for cell and gene therapies and hybrid artificial organs.

    DOI: 10.1097/01.MAT.0000093747.89681.4C

    Web of Science

    PubMed

    researchmap

  • BRPK, a novel protein kinase showing increased expression in mouse cancer cell lines with higher metastatic potential Reviewed

    A Nakajima, K Kataoka, M Hong, M Sakaguchi, N Huh

    CANCER LETTERS   201 ( 2 )   195 - 201   2003.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCI IRELAND LTD  

    A novel protein kinase named BRPK was isolated and partially characterized. BRPK was expressed at a higher level in three carcinoma cell lines with higher metastatic potential. Mouse and human BRPK cDNAs are well conserved and encode 580 and 581 amino acids, respectively. BRPK has a serine/threonine-type protein kinase domain, and the recombinant proteins of BRPK were capable of autophosphorylation. The results of a comparative sequence analysis indicated a possible link of BRPK to BRAP2. BRAP2 is known to bind the nuclear localization signal of BRCA1. We cloned mouse BRAP2 cDNA and showed the presence of isoforms. (C) 2003 Elsevier Ireland Ltd. All rights reserved.

    DOI: 10.1016/S0304-3835(03)00443-9

    Web of Science

    PubMed

    researchmap

  • Establishment of immortalized human hepatic stellate scavenger cells to develop bioartificial livers Reviewed

    T Watanabe, N Shibata, KA Westerman, T Okitsu, JE Allain, M Sakaguchi, T Totsugawa, M Maruyama, T Matsumura, H Noguchi, S Yamamoto, M Hikida, A Ohmori, M Reth, A Weber, N Tanaka, P Leboulch, N Kobayashi

    TRANSPLANTATION   75 ( 11 )   1873 - 1880   2003.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Background. Maintenance of liver-specific functions has been shown to be stabilized by co-cultivation of hepatocytes with hepatic stellate cells (HSC). Because the limited lifespan of human HSC is a major hurdle to their use, the authors report here the amplification of human HSC populations in vitro by retroviral transfer of human telomerase reverse transcriptase (hTERT).
    Methods. Human HSC strain LI 90 cells were transduced with a retroviral vector SSR#197 expressing hTERT and green fluorescent protein (GFP) cDNA flanked by a pair of loxP. TWNT-1, one of SSR#197-immortalized HSC, was characterized. Differentiated liver functions were evaluated in an immortalized human hepatocyte NKNT-3-TWNT-1 co-culture system.
    Results. TWNT-1 cells showed differential functions of HSC, including uptake of acetylated low-density lipoproteins and synthesis of collagen type I and hepatocyte growth factor. Efficient excision of the retro-virally transferred hTERT and GFP cDNAs was achieved by TAT-mediated expression of the Cre recombinase and subsequent GFP-negative cell sorting. When co-cultured with TWNT-1 cells, NKNT-3 increased protein expression of the detoxifying cytochrome P450-associated protein isoenzymes 3A4 and 2C9 and urea synthesis.
    Conclusions. TWNT-1 cells could be valuable in the study of integrated liver functions and contribute to the optimization of liver cell therapies and bioartificial livers.

    DOI: 10.1097/01.TP.0000064621.50907.A6

    Web of Science

    PubMed

    researchmap

  • Involvement of arachidonic acid in nonimmunologic production of superoxide in mast cells Reviewed

    M Sakaguchi, N Fukuishi, K Teramoto, M Miyazaki, NH Huh, M Namba, M Akagi

    INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY   130 ( 4 )   288 - 299   2003.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:KARGER  

    Background: A number of different molecules are known to be involved in the signal pathway to release histamine from mast cells, among which arachidonic acid (AA) is one of the key mediators. On the other hand, we found that the application of compound 48/80, a typical histamine liberator, generated superoxide in mast cells. In the present study, we investigated the mechanism of superoxide production in mast cells with respect to AA signaling in conjunction with a fine structural analysis. Methods: Superoxide production was monitored by chemiluminescence in rat peritoneal mast cells and their subfractions after various treatments. For scanning electron micrography, the conditions for fixation and freeze-fracture were optimized to get natural fine images. Results: Compound 48/80 induced superoxide production in the isolated mast cells and some of their subfractions possibly through intracellular increase in Ca2+ concentration, activation of cytosolic phospholipase A(2), and release of AA. Discussion: The present results indicate the critical role of AA in the signal pathway to generate superoxide from mast cells in response to compound 48/80. Copyright (C) 2003 S. Karger AG, Basel.

    DOI: 10.1159/000070216

    Web of Science

    researchmap

  • Active expression of p21 facilitates differentiation of immortalized human hepatocytes Reviewed

    N. Kobayashi, T. Kunieda, M. Sakaguchi, T. Okitsu, T. Totsugawa, M. Maruyama, Y. Kosaka, M. Takesue, N. Shibata, N. Tanaka

    Transplantation Proceedings   35 ( 1 )   433 - 434   2003

     More details

    Language:English   Publishing type:Research paper (international conference proceedings)   Publisher:Elsevier Inc.  

    DOI: 10.1016/S0041-1345(02)03784-3

    Scopus

    PubMed

    researchmap

  • Establishment of an immortalized human hepatic stellate cell line to develop antifibrotic therapies Reviewed

    Norikuni Shibata, Takamasa Watanabe, Teru Okitsu, Masakiyo Sakaguchi, Michihiko Takesue, Takemi Kunieda, Kenji Omoto, Shinichiro Yamamoto, Noriaki Tanaka, Naoya Kobayashi

    Cell Transplantation   12 ( 5 )   499 - 507   2003

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Cognizant Communication Corporation  

    Because human hepatic stellate cells (HSCs) perform a crucial role in the progress of hepatic fibrosis, it is of great value to establish an immortalized human cell line that exhibits HSC characteristics and grows well in tissue cultures for the development of antifibrotic therapies. Thus, we engineered an immortalized human hepatic stellate cell (HSC) line TWNT-4 by retrovirally inducing human telomerase reverse transcriptase (hTERT) into LI 90 cells established from a human liver mesenchymal tumor. Parental LI 90 entered replicative senescence, whereas TWNT-4 showed telomerase activity and proliferated for more than population doubling level (PDL) 200 without any crisis. TWNT-4 expressed platelet-derived growth factor-β receptor (PDGF-βR), α-smooth muscle actin (α-SMA), and type I collagen (α1) and was considered to be an activated form of HSCs. Treatment of TWNT-4 cells with either 100 U/ml of IFN-γ or 1 ng/ml of rapamycin (Rapa) for 14 days led to lower expression of type I collagen (α1) at RNA and protein levels. Exposure of TWNT-4 cells to both of IFN-γ (10 U/ml) and Rapa (0.1 ng/ml) for 14 days effectively decreased the expression of type I collagen (α1), PDGF-βR, and α-SMA expression and suppressed TGF-β1 secretion of TWNT-4 cells. We successfully induced apoptosis by transducing TNF-related apoptosis-inducing ligand (TRAIL) into TWNT-4 cells using adenovirus vectors Ad/GT-TRAIL and Ad/PGK-GV-17. These findings suggested that immortalized activated HSC line TWNT-4 would be a useful means to develop antifibrotic therapies.

    DOI: 10.3727/000000003108747064

    Scopus

    PubMed

    researchmap

  • Improved conditions to induce hepatocytes from rat bone marrow cells in culture Reviewed

    M Miyazaki, Akiyama, I, M Sakaguchi, E Nakashima, M Okada, K Kataoka, NH Huh

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   298 ( 1 )   24 - 30   2002.10

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Recent studies have revealed that bone marrow cells can develop into hepatocytes by in vivo transplantation under certain circumstances. However, little is known about the mechanism of bone marrow cell differentiation into hepatocytes. It is important to determine suitable culture conditions in which bone marrow cells will be differentiated into hepatocytes not only for understanding differentiation mechanisms but also for efficient amplification of hepatocyte-progenitor cells of bone marrow origin, this being a prerequisite for potential therapeutic use. In the present study, we found that hepatocyte growth factor (HGF) receptor (c-Met)- and a-fetoprotein-expressing cells were present in adult rat bone marrow. We also found that these cells also express hematopoietic stem cell markers, such as CD34, Thy-1, and c-Kit. Using an HGM medium with HGF and EGF, we succeeded in propagating hepatocyte-like cells induced from adult rat bone marrow in culture. These cells were immunocytochemically stained for albumin. By RT-PCR analysis of cultures containing the hepatocyte-like cells, we detected mRNAs of tryptophan-2,3-dioxygenase and tyrosine aminotransferase, markers of hepatocytes at a terminal differentiation stage. The present culture therefore can be a useful resource for cell transplantation therapy for liver diseases. (C) 2002 Elsevier Science (USA). All rights reserved.

    DOI: 10.1016/S0006-291X(02)02340-9

    Web of Science

    PubMed

    researchmap

  • Localization of S100C immunoreactivity in various human tissues Reviewed

    A Kondo, M Sakaguchi, E Makino, M Namba, S Okada, NH Huh

    ACTA MEDICA OKAYAMA   56 ( 1 )   31 - 34   2002.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:OKAYAMA UNIV MED SCHOOL  

    Using 2-dimensional gel electrophoresis, we previously demonstrated that the S100C protein remarkably decreased after immortalization of normal human fibroblasts, and that this protein caused growth inhibition of human tumor cells when forcibly expressed in these cells, suggesting that S100C plays a significant role in tumor suppression. The present study was carried out to determine what type of human tissues express S100C protein,, and, subsequently, whether the S100C content in these tissues changes after normal cells have been transformed into cancer cells. We found that ductal cells in various tissues were positively stained with the S100C protein. In comparison, epithelial cells in digestive organs such as, the stomach, small intestine, and colon were not stained as strongly. When 14 pairs of human normal and cancerous tissues were stained with the antibody, decreases in the staining levels of S100C were observed in 6 kinds of:cancerous tissues-from the bronchus, mammary duct, renal tubule, prostate, uterus, and testis-in comparison with staining in their normal counterparts. These results suggest that S100C is a new tumor marker protein, the expression of which significantly decreases after malignant transformation of human tissues.

    Web of Science

    researchmap

  • Antiproliferative activity of REIC/Dkk-3 and its significant down-regulation in non-small-cell lung carcinomas Reviewed

    T Tsuji, Nozaki, I, M Miyazaki, M Sakaguchi, H Pu, Y Hamazaki, O Iijima, M Namba

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   289 ( 1 )   257 - 263   2001.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC  

    We recently reported the cloning of the REIC/Dkk-3 gene, whose expression was shown to be downregulated in many human immortalized and tumor-derived cell lines [T. Tsuji et al. (2000) Biochem. Biophys. Res. Commun. 268, 20-24]. In the present study, we demonstrated that expression of the exogenous REIC/Dkk-3 gene in tumor cells inhibited cell growth. Furthermore, the level of REIC/Dkk-3 mRNA in normal human cells was lowest in the late G, phase during the cell cycle. Then we found that the expression of REIC/Dkk-3 was significantly down-regulated in surgically resected non-small-cell lung carcinomas. We determined the REIC/Dkk-3 locus on chromosome 11p15, where loss of heterozygosity has frequently been observed in human tumors. These findings indicate that REIC/Dkk-3 may function as a tumor suppressor. (C) 2001 Academic Press.

    Web of Science

    researchmap

  • Up-regulation of S100C in normal human fibroblasts in the process of aging in vitro Reviewed

    M Sakaguchi, M Miyazaki, T Kondo, M Namba

    EXPERIMENTAL GERONTOLOGY   36 ( 8 )   1317 - 1325   2001.8

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PERGAMON-ELSEVIER SCIENCE LTD  

    S100 proteins belonging to the EF-hand Ca2+ -binding protein family regulate a variety of cellular processes via interaction with different target proteins. Several diseases, including cancer and Alzheimer's disease, are related to a disorder of multifunctional S100 proteins, which are expressed in cell- and tissue-specific manners. We previously demonstrated that S100C could move to and accumulate in the nuclei of normal human fibroblasts but not in the nuclei of immortalized and neoplastic cells. In addition, we found that its nuclear accumulation resulted in suppression of DNA synthesis in normal cells at a confluent stage. In the present study, we investigated whether S100C was associated with cellular senescence in vitro. We found that S100C expression increased in normal human fibroblasts in the process of aging in culture and was accompanied by accumulation of its protein in the nuclei of senescent fibroblasts. In addition, the nuclear accumulation of S100C increased expression of a cyclin-dependent kinase inhibitor p21 (Sdil), a strong inhibitor of cell growth. These findings suggest that an increase in the cells having nuclear accumulation of S100C is closely related to the process of cellular senescence of normal human fibroblasts. (C) 2001 Elsevier Science Inc. All rights reserved.

    Web of Science

    researchmap

  • Establishment of a human hepatocyte line (OUMS-29) having CYP 1A1 and 1A2 activities from fetal liver tissue by transfection of SV-10 LT Reviewed

    KI Fukaya, S Asahi, S Nagamori, M Sakaguchi, C Gao, M Miyazaki, M Namba

    IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL   37 ( 5 )   266 - 269   2001.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:SOC IN VITRO BIOLOGY  

    Immortalized human hepatocytes that can retain functions of drug-metabolizing enzymes would be useful for medical and pharmacological studies and for constructing an artificial liver. The aim of this study wa. to establish immortalized human hepatocyte lines having differentiated liver-specific functions. pSVneo deoxyribonucleic acid, which contains large and small T genes in the early region of simian virus 40, was introduced into hepatocytes that had been obtained from the liver of a 21-wk-old fetus. Neomycin-resistant immortalized colonies were cloned and expanded to mass cultures to examine hepatic functions. Cells were cultured in a chemically defined serum-free medium, ASF101, which contains no peptides other than recombinant human transferrin and insulin. As a result, air immortal human hepatocyte cell line (OUMS-29) having li er-specific functions was established from one of the 13 clones. Expression of CYP 1A1 arid 1A2 messenger ribonucleic acid by the cells was induced by treatment with benz[a]pyrene, 3-methylcholanthrene, and benz[a]anthracene. OUMS-29 cells had both the polycyclic aromatic hydrocarbon receptor (AhR) arid AhR nuclear translocator. Consequently, 7-ethoxyresorufin deethylase activity of the cells was induced time- and dose-dependent by these polycyclic aromatic hydrocarbons. This cell line is expected to he instrumental as an alternative method in animal experiments for studying hepatocarcinogenesis. drug metabolisms of liver cells, and hepatic toxicology,.

    Web of Science

    researchmap

  • Overexpression of platelet-derived growth factor B and downregulation of PDGF-receptor alpha in human immortalized fibroblasts Reviewed

    C Gao, M Miyazaki, T Kondo, T Tsuji, M Sakaguchi, M Namba

    INTERNATIONAL JOURNAL OF ONCOLOGY   18 ( 4 )   871 - 875   2001.4

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PROFESSOR D A SPANDIDOS  

    Since immortalization of cells is critical in multistep carcinogenesis, efforts should be made to elucidate the mechanisms of the immortalization. To determine whether platelet-derived growth factor (PDGF) signal pathways play a role in immortalization of cells, we compared mRNA expressions of PDGFs and their receptors in three immortalized human fibroblast cell lines (SUSM-1, OUMS-24F, and KMST-6) with their normal parent cells. As a result, mRNA expression of PDGF-B (oncogene: c-sis) was upregulated in these immortalized cells. Unexpectedly, the expression of alpha- and beta -PDGF receptor genes was downregulated. PDGFR-alpha mRNA was remarkably decreased. When exogenous PDGFR-a was expressed transiently in the KMST-6 cells, the morphology of the cells resembled that of normal cells. These results suggest that the overexpression of PDGF-B (c-sis) and downregulation of PDGFR-alpha are related to the phenotypic characteristics of immortalized human cells.

    Web of Science

    researchmap

  • Two-dimensional gel electrophoretic studies on the cellular aging: accumulation of alpha-2-macroglobulin in human fibroblasts with aging

    T. Kondo, M. Sakaguchi, M. Namba

    Experimental Gerontology   36 ( 3 )   487 - 495   2001.3

     More details

    Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/s0531-5565(00)00256-4

    researchmap

  • Identification of a phosphoprotein that is downregulated in immortalized human fibroblasts Reviewed

    M Sakaguchi, M Miyazaki, T Kondo, T Tsuji, H Kouchi, M Namba

    ELECTROPHORESIS   22 ( 1 )   155 - 160   2001.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-V C H VERLAG GMBH  

    Many lines of evidence indicate that the immortalization step is critical for the neoplastic transformation of normal human cells. Once normal human cells have been immortalized, they are relatively easily transformed into neoplastic cells. In order to understand these phenomena, patterns of protein phosphorylation in proliferating normal human fibroblast cell strains and their immortalized cell lines were compared by using two-dimensional polyacrylamide gel electrophoresis. It was found that the expression and phosphorylation levels of the human heat shock protein 27 (HSP27) were predominantly downregulated in the immortalized cells compared with those in their normal counterparts. In the normal cells, HSP27 expression and phosphorylation were markedly increased by physiological and nonphysiological stresses, such as serum addition, treatment with a carcinogenic agent like 4-nitroquinoline-1-oxide, and a high osmotic pressure. This may be a normal defense against acute changes of cellular environment and cytotoxic effects. However, these stresses had no effects on the expression and phosphorylation of HSP27 in the immortalized cells. These results suggest that an abnormal regulation of HSP27 expression and phosphorylation may be one of the reasons for easy neoplastic transformation of the immortalized cells by the treatment with carcinogenic agents.

    Web of Science

    researchmap

  • Loss of nuclear localization of the S100C protein in immorialized human fibroblasts Reviewed

    M Sakaguchi, H Yamada, T Tsuji, Y Inoue, M Miyazaki, T Tanaka, M Namba

    RADIATION RESEARCH   155 ( 1 )   208 - 214   2001.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:RADIATION RESEARCH SOC  

    It is well known that cancer develops through a multistep process, lit vitro transformation studies of normal human cells have shown that the immortalization step is critical for neoplastic transformation of cells. Furthermore, studies of cell fusion between normal and immortalized cells have indicated that the normal phenotype is dominant and the immortal phenotype is recessive. Thus we looked for cellular proteins that were down-regulated in immortalized human cells by two-dimensional gel electrophoresis to elucidate the mechanisms of immortalization of human cells. We found that the S100C protein was down-regulated in immortalized cells. This protein was localized in the cytoplasm of cells at the semiconfluent stage, while at the confluent stage it moved into the nuclei of normal cells but not into those of immortalized cells. Microinjection of an S100C antibody into normal confluent cells diminished the level of nuclear S100C protein, resulting in DNA synthesis. Taken together, loss of nuclear localization of the S100C protein, which may be related to DNA synthesis, is thought to be one of the mechanisms of immortalization, (C) 2001 by Radiation Research Society.

    Web of Science

    researchmap

  • Hepatocyte growth factor induces differentiation of adult rat bone marrow cells into a hepatocyte lineage in vitro Reviewed

    SH Oh, M Miyazaki, H Kouchi, Y Inoue, M Sakaguchi, T Tsuji, N Shima, K Higashio, M Namba

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   279 ( 2 )   500 - 504   2000.12

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC  

    Bone marrow (BM) cells originally include alpha-fetoprotein (AFP)- and c-Met [a receptor for hepatocyte growth factor (HGF)]-expressing cells. In vitro treatment of BM cells with HGF induced albumin-expressing hepatocyte-like cells. Furthermore, those hepatocyte-like cells expressed cytokeratins 8 and 18, which are typically expressed in normal adult hepatocytes. These findings demonstrate that BM cells include AFP-expressing hepatic progenitor cells that can be differentiated into hepatocytes by HGF in culture, indicating that such cultures are useful resources for cell transplantation therapy for liver diseases. (C) 2000 Academic Press.

    DOI: 10.1006/bbrc.2000.3985

    Web of Science

    researchmap

  • Treatment of surgically induced acute liver failure with transplantation of highly differentiated immortalized human hepatocytes Reviewed

    N Kobayashi, M Miyazaki, K Fukaya, Y Inoue, M Sakaguchi, H Noguchi, T Matsumura, T Watanabe, T Totsugawa, N Tanaka, M Namba

    CELL TRANSPLANTATION   9 ( 5 )   733 - 735   2000.9

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:COGNIZANT COMMUNICATION CORP  

    Primary human hepatocytes are an ideal source of hepatic function in bioartficial liver (BAL), but the shortage of human livers available for hepatocyte isolation limits this modality. To resolve this issue, primary human fetal hepatocytes were immortalized using simian virus 40 large T antigen. One of the immortal cell lines, OUMS-29, showed highly differentiated liver functions. Intrasplenic transplantation of OUMS-29 cells protected 90% hepatectomized rats from hyperammonemia and significantly prolonged their survival. Essentially unlimited availability of OUMS-29 cells supports their clinical use for BAL treatment.

    Web of Science

    researchmap

  • Relationship between contact inhibition and intranuclear S100C of normal human fibroblasts Reviewed

    M Sakaguchi, M Miyazaki, Y Inoue, T Tsuji, H Kouchi, T Tanaka, H Yamada, M Namba

    JOURNAL OF CELL BIOLOGY   149 ( 6 )   1193 - 1206   2000.6

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ROCKEFELLER UNIV PRESS  

    Many lines of evidence indicate that neoplastic transformation of cells occurs by a multistep process. For neoplastic transformation of normal human cells, they must be first immortalized and then be converted into neoplastic cells. It is well known that the immortalization is a critical step for the neoplastic transformation of cells and that the immortal phenotype is recessive. Thus, we investigated proteins downregulated in immortalized cells by two-dimensional gel electrophoresis. As a result, S100C, a Ca2+-binding protein, was dramatically downregulated in immortalized human fibroblasts compared with their normal counterparts. When the cells reached confluence, S100C was phosphorylated on threonine 10. Then the phosphorylated S100C moved to and accumulated in the nuclei of normal cells, whereas in immortalized cells it was not phosphorylated and remained in the cytoplasm. Microinjection of the anti-S100C antibody into normal confluent quiescent cells induced DNA synthesis. Furthermore, when exogenous S100C was compelled to localize in the nuclei of HeLa cells, their DNA synthesis was remarkably inhibited with increase in cyclin-dependent kinase inhibitors such as p16(Ink4a) and p21(Waf1) These data indicate the possible involvement of nuclear S100C in the contact inhibition of cell growth.

    Web of Science

    researchmap

  • In vitro aging research in Japan Reviewed

    T Tsuji, M Miyazaki, M Sakaguchi, M Namba

    EXPERIMENTAL GERONTOLOGY   35 ( 3 )   291 - 298   2000.5

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:PERGAMON-ELSEVIER SCIENCE LTD  

    Web of Science

    researchmap

  • Establishment of a highly differentiated immortalized human hepatocyte cell line as a source of hepatic function in the bioartificial liver Reviewed

    N Kobayashi, M Miyazaki, K Fukaya, Y Inoue, M Sakaguchi, H Noguchi, N Tanaka, M Namba

    TRANSPLANTATION PROCEEDINGS   32 ( 2 )   237 - 241   2000.3

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ELSEVIER SCIENCE INC  

    Web of Science

    researchmap

  • Prevention of acute liver failure in rats with reversibly immortalized human hepatocytes Reviewed

    N Kobayashi, T Fujiwara, KA Westerman, Y Inoue, M Sakaguchi, H Noguchi, M Miyazaki, J Cai, N Tanaka, IJ Fox, P Leboulch

    SCIENCE   287 ( 5456 )   1258 - 1262   2000.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:AMER ASSOC ADVANCEMENT SCIENCE  

    Because of a critical shortage in suitable organs, many patients with terminal liver disease die each year before liver transplantation can be performed. Transplantation of isolated hepatocytes has been proposed for the temporary metabolic support of patients awaiting liver transplantation or spontaneous reversion of their liver disease. A major limitation of this form of therapy is the present inability to isolate an adequate number of transplantable hepatocytes. A highly differentiated cell line, NKNT-3, was generated by retroviral transfer in normal primary adult human hepatocytes of an immortalizing gene that can be subsequently and completely excised by Cre/Lox site-specific recombination. When transplanted into the spleen of rats under transient immunosuppression, reversibly immortalized NKNT-3 cells provided life-saving metabolic support during acute liver failure induced by 90% hepatectomy.

    Web of Science

    researchmap

  • A REIC gene shows down-regulation in human immortalized cells and human tumor-derived cell lines Reviewed

    T Tsuji, M Miyazaki, M Sakaguchi, Y Inoue, M Namba

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   268 ( 1 )   20 - 24   2000.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC  

    Normal human cells stop proliferation after a certain number of cell divisions. This phenomenon is called cellular aging. The fact that the senescence phenotype is dominant and the immortal one is recessive indicates that immortalization of human cells may be caused by loss of functions of certain genes in normal cells. Based on this evidence, several cDNA clones whose expression was down-regulated during the immortalization process of human cells were isolated by the representative difference analysis (RDA) system in our laboratory. One of them, which was named REIC, was expressed to a lower degree in three human immortalized cell lines as compared with their parental normal counterparts, In addition, the expression of REIC was markedly lower in eight human tumor-derived cell lines (Hep3B and HuH-7 hepatocellular carcinomas, HuH-6 Clone 5 hepatoblastoma, HuCCT-1 cholangiocarcinoma, A549 lung cancer, HaCaT immortalized keratinocyte, HeLa cervical carcinoma, and Saos-2 osteosarcoma). In contrast, among the human tissues examined, the heart and brain, which contain a large number of post-mitotic cells, showed the highest expression of REIC. The full-length REIC cDNA revealed that the predicted protein is 350 amino acids in length and possesses coiled-coil tertiary structures in each of the amino- and carboxyl-termini. Furthermore, a search of the protein database revealed a match of this gene product with Dkk-3, which is a novel inhibitor of Wnt oncogene. These results indicate that the REIC cloned by us may function as a tumor suppressor. (C) 2000 Academic Press.

    Web of Science

    researchmap

  • Transplantation of highly differentiated immortalized human hepatocytes to treat acute liver failure Reviewed

    N Kobayashi, M Miyazaki, K Fukaya, Y Inoue, M Sakaguchi, T Uemura, H Noguchi, A Kondo, N Tanaka, M Namba

    TRANSPLANTATION   69 ( 2 )   202 - 207   2000.1

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Background. Temporary support of a damaged liver by a bioartificial liver (BAL) devise is a promising approach for the treatment of acute liver failure. Although human primary hepatocytes are an ideal source of hepatic function in BAL, shortage of human livers available for hepatocyte isolation is the limiting factor for the use of this modality. A clonal human hepatocyte cell line that can grow economically in culture and exhibit liver-specific functions should be an attractive solution to this problem.
    Methods. To test this alternative, primary human fetal hepatocytes were immortalized using Simian virus 40 large T antigen. To investigate the potential of the immortalized cells for BAL, we transplanted the cells into the spleen of adult rats and performed a 90% hepatectomy 12 hr later.
    Results, One of the cloned human liver cell lines, OUMS-29, showed highly differentiated liver functions. Intrasplenic transplanting of 20x10(6) OUMS-29 cells protected the animals from hyperammonemia and the associated hepatic encephalopathy, Survival was significantly prolonged in 90% of hepatectomized rats receiving GUMS-29 cells.
    Conclusions. A highly differentiated immortalized human hepatocyte cell line, OUMS-29, was able to provide metabolic support during acute liver failure induced by 90% hepatectomy in rats. Essentially unlimited availability of OUMS-29 cells may be clinically useful for BAL treatment.

    Web of Science

    PubMed

    researchmap

  • Establishment of a human hepatoma cell line, HLE/2E1, suitable for detection of P450 2E1-related cytotoxicity Reviewed

    Isao Nozaki, Toshiya Tsuji, Masakiyo Sakaguchi, Yusuke Inoue, Ryuji Hirai, Akio Andou, Masahiro Miyazaki, Nobuyoshi Shimizu, Masayoshi Namba

    In Vitro Cellular and Developmental Biology - Animal   36 ( 9 )   566 - 570   2000

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    By transfection of an expression vector of human cytochrome P450 2E1 (CYP2E1) into a human hepatoma cell line (HLE), a new cell line (HLE/2E1) that stably expresses activity of CYP2E1 has been established. The HLE/2E1 cell line expressed a higher level of CYP2E1 messenger ribonucleic acid than did the mother HLE cell line. CYP2E1 enzyme activity determined by a p-nitrophenol oxidation assay was also higher in HLE/2E1 cells than in HLE cells. In addition, the enzyme activity of the HLE/2E1 cells was increased by ethanol treatment. Exposure to acetaminophen (APAP) or buthionine sulfoximine (BSO) caused a greater decrease in viability of the HLE/2E1 cells than that of the HLE cells, as determined by the 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide assay. The cytotoxicity of APAP or BSO to HLE/2E1 cells was inhibited by the addition of ethanol or vitamin E. However, the cytotoxicity of both APAP and BSO was enhanced by 24-h preincubation of HLE/2E1 cells with ethanol. These results show that this cell line provides a useful model for studying catalytic properties of CYP2E1 and cytotoxic mechanisms of chemicals metabolized by CYP2E1.

    DOI: 10.1007/BF02577524

    Scopus

    PubMed

    researchmap

  • Manumycin A, inhibitor of ras farnesyltransferase, inhibits proliferation and migration of rat vascular smooth muscle cells Reviewed

    H Kouchi, K Nakamura, K Fushimi, M Sakaguchi, M Miyazaki, T Ohe, M Namba

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   264 ( 3 )   915 - 920   1999.11

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:ACADEMIC PRESS INC  

    Restenosis after angioplasty is thought to be caused by proliferation and migration of vascular smooth muscle cells (VSMCs), and it is a most serious problem in medical treatment. A low dose (50 ng/ml) of manumycin A, an inhibitor of p21(ras) (ras) farnesylation, significantly inhibited proliferation of rat VSMCs stimulated by the platelet-derived growth factor (PDGF). The mitoinhibitory effect of manumycin A was dose- and time-dependent but was independent of cell density. Western blot analysis showed that manumycin A reduced the amount of functional ras localized at the cytoplasmic membrane and inhibited the phosphorylation of p42/44 mitogen-activated protein kinase (MAPK). Manumycin A also inhibited VSMC migration and disorganized alpha actin fibers, as shown by immnofluorecence staining. These results indicate that the interruption of the ras/MAPK signal transduction pathway and the disorganization of alpha actin fibers are the main cause of manumycin A inhibition of VSMC proliferation and migration induced by PDGF. (C) 1999 Academic Press.

    Web of Science

    researchmap

  • Transferrin synthesized in cultured human fibroblasts is associated with tubulins and has iron binding capacity Reviewed

    Masakiyo Sakaguchi, Tadashi Kondo, Hong Pu, Masayoshi Namba

    Cell Structure and Function   24 ( 1 )   5 - 9   1999

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    In a previous report, using immunocytochemical and fluorescence-labeling techniques, we demonstrated that transferrin is synthesized in cultured human fibroblasts and that it is associated with tubulins in the cells. These morphological findings led us to attempt to elaborate those issues in more detail by biochemical methods. In this report, we were able to prove the association of transferrin produced in cells with tubulins. In addition, the transferrin associated with tubulins was found to bind to iron. These results suggest that endogenous transferrin plays a role in preventing damage caused by free radicals which can be induced by the interaction of iron with the hydrogen peroxide produced in cells.

    DOI: 10.1247/csf.24.5

    Scopus

    PubMed

    researchmap

  • Characteristics of intracellular transferrin produced by human fibroblasts: Its posttranscriptional regulation and association with tubulin Reviewed

    Tadashi Kondo, Masakiyo Sakaguchi, Masayoshi Namba

    Experimental Cell Research   242 ( 1 )   38 - 44   1998.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Academic Press Inc.  

    Transferrin (Tf), an iron-binding protein, was investigated in the cultured human fibroblast which is a major cell type in connective tissues. Tf is a major iron-transporting protein and has an important role in iron metabolism. Using two-dimensional gel electrophoresis, we demonstrated the eight subtypes of Tf produced by cultured normal human fibroblasts and their down-regulation after the immortalization of human cells, an essential early step of in vitro transformation. However, the amount of Tf mRNA in the immortalized cells was equal to that in the normal human fibroblasts, suggesting that the down-regulation occurred at the posttranscriptional level. The amount of Tf receptor increased in the immortalized cells in spite of a decrease in the amount of intracellular Tf. Interestingly the produced Tf associated with microtubules. These findings suggest a novel aspect of Tf characteristics in human fibroblasts.

    DOI: 10.1006/excr.1998.4080

    Scopus

    PubMed

    researchmap

  • Two-dimensional gel electrophoretic analysis of the changes after immortalization of human cells: Decrease of intracellular alpha-2-macroglobulin fragment Reviewed

    T Kondo, M Sakaguchi, H Yamada, M Namba

    ELECTROPHORESIS   19 ( 10 )   1836 - 1840   1998.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:WILEY-V C H VERLAG GMBH  

    To study the mechanisms of immortalization of human cells, an early step in cancer development, we compared the cellular proteins of normal and immortalized human fibroblasts. Two-dimensional gel electrophoresis showed that one spot with a molecular mass 20 kDa and an isoelectric point of 6.0, became significantly smaller after immortalization of human cells. Further, the spot was rarely observed in four human liver cancer cell lines. Investigation of the N-terminal amino acids revealed that the spot was a fragment of alpha-2-macroglobulin. Although the 20 kDa fragment contains methionine, the spot was not labeled with [S-35]methionine. Thus we concluded that the spot might be derived from the culture medium. These results indicated that intracellular metabolism of alpha-2-macroglobulin, which is a multifunctional protease inhibitor, changed after the cells were transformed.

    Web of Science

    researchmap

  • Differential localization of two types of transferrin: Produced by human fibroblasts or incorporated from culture medium Reviewed

    Masakiyo Sakaguchi, Tadashi Kondo, Hong Pu, Masayoshi Namba

    Cell Structure and Function   23 ( 2 )   69 - 72   1998

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Japan Society for Cell Biology  

    In a previous paper, we demonstrated that cultured human fibroblasts synthesize transferrin (Tf). Two types of Tf are present
    one is produced by the cells and the other is internalized from the culture medium. To study the metabolism of intracellular Tf, we investigated the subcellular localization of the two types of Tf in human fibroblasts by immunocytochemical and fluorescence-labeling techniques. The internalized Tf was found to be localized in the perinuclear area, and the synthesized Tf was associated with microtubules, forming a fibrous structure in the cytoplasm. When the cells were treated with colchicine which depolymerizes microtubules irreversibly, the synthesized Tf lost its fibrous structure and spread out in cytoplasm, but the internalized Tf remained around the nucleus. These results suggest that the two types of Tf are regulated differently in the cells.

    DOI: 10.1247/csf.23.69

    Scopus

    PubMed

    researchmap

▼display all

Books

  • 動物細胞培養・自動化におけるトラブル発生原因と対策

    技術情報協会  2017 

     More details

  • 培養細胞実験ハンドブック

    羊土社  2009 

     More details

  • 培養細胞実験ハンドブック

    羊土社  2009 

     More details

  • 培養細胞実験ハンドブック

    羊土社  2009 

     More details

  • 培養細胞実験ハンドブック

    羊土社  2004 

     More details

  • 培養細胞実験ハンドブック

    羊土社  2004 

     More details

  • 培養細胞実験ハンドブック

    羊土社  2004 

     More details

▼display all

MISC

  • S100A11は筋浸潤性膀胱癌細胞と線維芽細胞のクロストークに関与し腫瘍進行に寄与する(S100A11 contributes to tumor progression with cross talking between muscle invasive bladder cancer cells and fibroblasts)

    光井 洋介, 定平 卓也, 渡部 昌実, 丸山 雄樹, 荒木 元朗, 渡邉 豊彦, 阪口 政清, 那須 保友

    西日本泌尿器科   81 ( 増刊 )   138 - 138   2019.10

     More details

    Language:English   Publisher:西日本泌尿器科学会  

    researchmap

  • S100タンパク質に着眼したがん転移機構の解明とその制御.

    SAKAGUCHI Masakiyo

    岡山医学会雑誌   130   135 - 139   2018.10

     More details

    Language:Japanese  

    researchmap

  • 膵がん進展に導く膵がん細胞 間質線維芽細胞クロストークを介在する分泌性S100A11 受容体RAGE連携の役割

    光井 洋介, 山本 健一, Sumardika I Wayan, 木下 理恵, 村田 等, 二見 淳一郎, 高松 仁, 山本 靖彦, 西堀 正洋, 豊岡 伸一, 渡部 昌実, 那須 保友, 阪口 政清

    日本がん免疫学会総会プログラム・抄録集   22回   156 - 156   2018.7

     More details

    Language:Japanese   Publisher:日本がん免疫学会  

    researchmap

  • 分泌性S100A11-受容体RAGEシグナルに着眼した膵がん間質増大のメカニズムの解明

    山本 健一, 高松 仁, 光井 洋介, 木下 理恵, 村田 等, 二見 淳一郎, 山本 靖彦, 西堀 正洋, 豊岡 伸一, 阪口 政清

    日本がん免疫学会総会プログラム・抄録集   22回   117 - 117   2018.7

     More details

    Language:Japanese   Publisher:日本がん免疫学会  

    researchmap

  • Signal Diversity of Receptor for Advanced Glycation End Products

    Masakiyo Sakaguchi, Rie Kinoshita, Endy Widya Putranto, I. Made Winarsa Ruma, I. Wayan Sumardika, Chen Youyi, Naoko Tomonobu, Ken-ichi Yamamoto, Hitoshi Murata

    ACTA MEDICA OKAYAMA   71 ( 6 )   459 - 465   2017.12

     More details

    Language:English   Publishing type:Book review, literature introduction, etc.   Publisher:OKAYAMA UNIV MED SCHOOL  

    The receptor for advanced glycation end products (RAGE) is involved in inflammatory pathogenesis. It functions as a receptor to multiple ligands such as AGEs, HMGB1 and S100 proteins, activating multiple intracellular signaling pathways with each ligand binding. The molecular events by which ligand-activated RAGE controls diverse signaling are not well understood, but some progress was made recently. Accumulating evidence revealed that RAGE has multiple binding partners within the cytoplasm and on the plasma membrane. It was first pointed out in 2008 that RAGE's cytoplasmic tail is able to recruit Diaphanous-1 (Dia-1), resulting in the acquisition of increased cellular motility through Rac1/Cdc42 activation. We also observed that within the cytosol, RAGE's cytoplasmic tail behaves similarly to a Toll-like receptor (TLR4)-TIR domain, interacting with TIRAP and MyD88 adaptor molecules that in turn activate multiple downstream signals. Subsequent studies demonstrated the presence of an alternative adaptor molecule, DAP10, on the plasma membrane. The coupling of RAGE with DAP10 is critical for enhancing the RAGE-mediated survival signal. Interestingly, RAGE interaction on the membrane was not restricted to DAP10 alone. The chemotactic G-protein-coupled receptors (GPCRs) formyl peptide receptors1 and 2 (FPR1 and FPR2) also interacted with RAGE on the plasma membrane. Binding interaction between leukotriene B4 receptor 1 (BLT1) and RAGE was also demonstrated. All of the interactions affected the RAGE signal polarity. These findings indicate that functional interactions between RAGE and various molecules within the cytoplasmic area or on the membrane area coordinately regulate multiple ligand-mediated RAGE responses, leading to typical cellular phenotypes in several pathological settings. Here we review RAGE's signaling diversity, to contribute to the understanding of the elaborate functions of RAGE in physiological and pathological contexts.

    Web of Science

    researchmap

  • Exogenous DKK-3/REIC inhibits Wnt/beta-catenin signaling and cell proliferation in human kidney cancer KPK1

    Jiaqi Xu, Takuya Sadahira, Rie Kinoshita, Shun-Ai Li, Peng Huang, Koichiro Wada, Motoo Araki, Kazuhiko Ochiai, Hirofumi Noguchi, Masakiyo Sakaguchi, Yasutomo Nasu, Masami Watanabe

    ONCOLOGY LETTERS   14 ( 5 )   5638 - 5642   2017.11

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    The third member of the Dickkopf family (DKK-3), also known as reduced expression in immortalized cells (REIC), is a tumor suppressor present in a variety of tumor cells. Regarding the regulation of the Wnt/beta-catenin signaling pathway, exogenous DKK-1 and DKK-2 are reported to inhibit Wnt signaling by binding the associated effectors. However, whether exogenous DKK-3 inhibits Wnt signaling remains unclear. A recombinant protein of human full-length DKK-3 was used to investigate the exogenous effects of the protein in vitro in KPK1 human renal cell carcinoma cells. It was demonstrated that the expression of phosphorylated (p-)beta-catenin (inactive form as the transcriptional factor) was increased in KPK1 cells treated with the exogenous DKK-3 protein. The levels of non-p-beta-catenin (activated form of beta- catenin) were consistently decreased. It was revealed that the expression of transcription factor (TCF) 1 and c-Myc, the downstream transcription factors of the Wnt/beta-catenin signaling pathway, was inhibited following treatment with DKK-3. A cancer cell viability assay confirmed the anti-proliferative effects of exogenous DKK-3 protein, which was consistent with a suppressed Wnt/beta-catenin signaling cascade. In addition, as low-density lipoprotein receptor-related protein 6 (LRP6) is a receptor of DKK-1 and DKK-2 and their interaction on the cell surface inhibits Wnt/beta-catenin signaling, it was examined whether the exogenous DKK-3 protein affects LRP6-mediated Wnt/beta-catenin signaling. The LRP6 gene was silenced and the effects of DKK-3 on the time course of the upregulation of p-beta-catenin expression were subsequently analyzed. Notably, LRP6 depletion elevated the base level of p-beta-catenin; however, there was no significant effect on its upregulation course or expression pattern. These findings indicate that exogenous DKK-3 upregulates p-beta-catenin and inhibits Wnt/beta-catenin signaling in an LRP6-independent manner. Therefore, exogenous DKK-3 protein may inhibit the proliferation of KPK1 cells via inactivating Wnt/beta-catenin signaling.

    DOI: 10.3892/ol.2017.6833

    Web of Science

    researchmap

  • Promising therapeutic efficacy of a novel reduced expression in immortalized cells/dickkopf-3 expressing adenoviral vector for hepatocellular carcinoma

    Hiroaki Sawahara, Hidenori Shiraha, Daisuke Uchida, Hironari Kato, Ryo Kato, Atsushi Oyama, Teruya Nagahara, Masaya Iwamuro, Shigeru Horiguchi, Koichiro Tsutsumi, Mari Mandai, Tetsushige Mimura, Nozomu Wada, Yasuto Takeuchi, Kenji Kuwaki, Hideki Onishi, Shinichiro Nakamura, Masami Watanabe, Masakiyo Sakaguchi, Akinobu Takaki, Kazuhiro Nouso, Takahito Yagi, Yasutomo Nasu, Hiromi Kumon, Hiroyuki Okada

    JOURNAL OF GASTROENTEROLOGY AND HEPATOLOGY   32 ( 10 )   1769 - 1777   2017.10

     More details

    Language:English   Publisher:WILEY  

    Background and Aim: Reduced expression in immortalized cells (REIC)/dickkopf-3 (Dkk-3) is a tumor suppressor gene that is downregulated in various cancers. In our previous study of prostate cancer, the REIC/Dkk-3-expressing adenoviral vector (Ad-REIC) was found to induce cancer-selective apoptosis. This study recently developed a novel super gene expression (SGE) system and used this system to re-construct an Ad-REIC vector, termed the Ad-SGE-REIC, to achieve more effective therapeutic outcomes. In this study, the therapeutic effects of Ad-SGE-REIC on hepatocellular carcinoma (HCC) was assessed.
    Methods: Human HCC cell lines (HLE, Huh7, HepG2, HLF, SK-Hep1, and PLC), human HCC tissues, and mouse HCC cell line (Hepa1-6) were used in this study. REIC/Dkk-3 expression was assessed by immunoblotting and immunohistochemistry. The relative cell viability and the apoptotic effect were examined in vitro, and the anti-tumor effects of Ad-SGE-REIC treatment were analyzed in the mouse xenograft model. This study additionally assessed anti-tumor immunological effects on the immunocompetent mice.
    Results: REIC/Dkk-3 expression was decreased in HCC cell lines and HCC tissues. Ad-SGE-REIC reduced cell viability and induced apoptosis in HCC cell lines (HLE and Huh7), inhibited tumor growth in the mouse xenograft model, and demonstrated in vivo anti-cancer immunostimulatory effects on the HCC cell line (Hepa1-6).
    Conclusions: Ad-SGE-REIC treatment not only enhanced cell killing effects in vitro but also elicited significant therapeutic effects, with tumor growth suppression, in vivo. REIC/Dkk-3 gene therapy using Ad-SGE-REIC potentially represents an innovative new therapeutic tool for HCC.

    DOI: 10.1111/jgh.13757

    Web of Science

    researchmap

  • Identification of a novel component leading to anti-tumor activity besides the major ingredient cordycepin in Cordyceps militaris extract

    Takeharu Wada, I. Wayan Sumardika, Shingo Saito, I. Made Winarsa Ruma, Eisaku Kondo, Masami Shibukawa, Masakiyo Sakaguchi

    JOURNAL OF CHROMATOGRAPHY B-ANALYTICAL TECHNOLOGIES IN THE BIOMEDICAL AND LIFE SCIENCES   1061   209 - 219   2017.9

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    In accordance with our previous study that was carried out to identify novel anti-tumor ingredients, chromatographic separation in combination with an anti-tumor activity assay was used for analysis of Cordyceps militaris extract in this study. Various modes of chromatography including reversed-phase, cation-exchange and anion exchange were used to separate components of Cordyceps militaris, which showed various chemical properties. Anti-tumor activity of each fraction was assessed by a Hoechst staining-based apoptosis assay using malignant melanoma MeWo cells. By these repeated approaches through chromatographic segregation and cell biological assay, we finally succeeded in identifying the target substance from a certain fraction that included neutral hydrophilic components using a pre-column and post-column chlorine adduct ionization LC APCI MS method. The target substance was a mono-carbohydrate, xylitol, that induced apoptotic cell death in MeWo cells but not in normal human OUMS-24 fibroblasts. This is the first study showing that Cordyceps militaris extract contains a large amount of xylitol. Thus, our results will contribute greatly to uncovering the mysterious multifunctional herbal drug Cordyceps militaris as an anti-tumor agent.

    DOI: 10.1016/j.jchromb.2017.07.022

    Web of Science

    researchmap

  • Robust cancer-specific gene expression by a novel cassette with hTERT and CMV promoter elements

    Masakiyo Sakaguchi, Takuya Sadahira, Hideo Ueki, Rie Kinoshita, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Yasutomo Nasu, Kazuhiko Ochiai, Hiromi Kumon, Nam-Ho Huh, Masami Watanabe

    ONCOLOGY REPORTS   38 ( 2 )   1108 - 1114   2017.8

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    We developed and validated a novel hTERT/CMV promoter element-driven gene expression cassette that can robustly enhance cancer-specific gene expression. The following gene expressional elements were located in tandem within the plasmid construct: [hTERT core promoter, cytomegalovirus (CMV) minimized promoter, RU5' sequence, an inserted gene, BGH polyA, hTERT enhancer]; this is hereafter referred to as the hT/Cm-R-hT construct. Using various human cancer cell lines and normal cells, the cancer-specific transcription of the green fluorescent protein (GFP) gene was examined by western blotting and fluorescence microscopy. Cancer-specific gene expression was robustly achieved in the hT/Cm-R-hT plasmid in comparison to the other control hT/Cm-driven construct. Notably, the expression level of GFP observed in the hT/Cm-RhT-driven construct was superior to that of the control plasmid with the conventional CMV promoter in HEK293 cells, which are known to possess higher hTERT activity than normal cells. We next examined the availability of hT/Cm-R-hT in detecting the target GFP expressing cancer cells from human peripheral blood mononuclear cells (PBMCs). The hT/Cm-R-hT plasmid successfully induced cancer-specific gene expression; the robust expression of GFP was observed in target HeLa cancer cells, whereas GFP was not visibly expressed in normal PBMCs. The plasmid allowed for the selective visualization of viable HeLa cancer cells in mixed cell cultures containing up to 10000-fold more PBMCs. These findings indicate that the hT/Cm-R-hT expressional system is a valuable tool for detecting viable cancer cells mixed with normal cells. The current system can therefore be applied to the in vitro detection of cancer cells that are disseminated in the blood and other types of body fluid in vivo. Since the current system can also be applied to other types of vectors, including virus vectors, this approach using the hTERT promoter-based construct is expected to become a valuable tool for enhancing cancer specific gene expression.

    DOI: 10.3892/or.2017.5710

    Web of Science

    researchmap

  • Expression of tumor suppressor REIC/Dkk-3 by a newly improved adenovirus vector with insertion of a hTERT promoter at the 3'-side of the transgene

    Endy Widya Putranto, Rie Kinoshita, Masami Watanabe, Takuya Sadahira, Hitoshi Murata, Ken-Ichi Yamamoto, Junichiro Futami, Ken Kataoka, Yusuke Inoue, I. Made Winarsa Ruma, I. Wayan Sumardika, Chen Youyi, Miyoko Kubo, Yoshihiko Sakaguchi, Kenji Saito, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh, Masakiyo Sakaguchi

    ONCOLOGY LETTERS   14 ( 1 )   1041 - 1048   2017.7

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Reduced expression in immortalized cells (REIC)/Dickkopf-3 (Dkk-3) overexpression, induced using an adenovirus (Ad)-REIC, has been revealed to have a dramatic therapeutic effect on multiple types of cancer. To achieve an improved therapeutic effect from Ad-REIC on cancer, our group previously developed an enhanced gene expression system, the C-TSC cassette [cytomegalovirus (CMV)-RU5' located upstream (C); another promoter unit composed of triple tandem promoters, human telomerase reverse transcriptase (hTERT), simian virus 40 and CMV, located downstream of the cDNA (TSC); plus a polyadenylation (polyA) signal]. When applied to the conventional Ad-REIC, this novel system induced the development of an enhanced product, Ad-C-TSC-REIC, which exhibited a noticeable anticancer effect. However, there were difficulties in terms of Ad-C-TSC-REIC productivity in HEK293 cells, which are a widely used donor cell line for viral production. Productivity of Ad-C-TSC-REIC was significantly reduced compared with the conventional Ad-REIC, as the Ad-C-TSC-REIC had a significantly higher ability to induce apoptotic cell death of not only various types of cancer cell, but also HEK293 cells. The present study aimed to overcome this problem by modifying the C-TSC structure, resulting in an improved candidate: A C-T cassette (C: CMV-RU5' located upstream; T: another promoter unit composed of a single hTERT promoter, located downstream of the cDNA plus a polyA signal), which demonstrated gene expression comparable to that of the C-TSC system. The improved adenovirus REIC/Dkk-3 product with the C-T cassette, named Ad-C-T-REIC, exhibited a higher expression level of REIC/Dkk3, similar to that of Ad-C-TSC-REIC. Notably, the vector mitigated the cell death of donor HEK293 cells, resulting in a higher rate of production of its adenovirus. These results indicated that Ad-C-T-REIC has the potential to be a useful tool for application in cancer gene therapy.

    DOI: 10.3892/ol.2017.6201

    Web of Science

    researchmap

  • An HNF4-microRNA-194/192 signaling axis maintains hepatic cell function

    Aoi Morimoto, Mana Kannari, Yuichi Tsuchida, Shota Sasaki, Chinatsu Saito, Tsuyoshi Matsuta, Tsukasa Maeda, Megumi Akiyama, Takahiro Nakamura, Masakiyo Sakaguchi, Nobukazu Nameki, Frank J. Gonzalez, Yusuke Inoue

    JOURNAL OF BIOLOGICAL CHEMISTRY   292 ( 25 )   10574 - 10585   2017.6

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Hepatocyte nuclear factor 4 (HNF4) controls the expression of liver-specific protein-coding genes. However, some microRNAs are also modulated by HNF4, and it is not known whether they are direct targets of HNF4 and whether they influence hepatic function. In this study, we found that HNF4 regulates microRNAs, indicated by marked down-regulation of miR-194 and miR-192 (miR-194/192) in liver-specific Hnf4a-null (Hnf4a(H)) mice. Transactivation of the shared miR-194/192 promoter was dependent on HNF4 expression, indicating that miR-194/192 is a target gene of HNF4. Screening of potential mRNAs targeted by miR-194/192 revealed that expression of genes involved in glucose metabolism (glycogenin 1 (Gyg1)), cell adhesion and migration (activated leukocyte cell adhesion molecule (Alcam)), tumorigenesis and tumor progression (Rap2b and epiregulin (Ereg)), protein SUMOylation (Sumo2), epigenetic regulation (Setd5 and Cullin 4B (Cln4b)), and the epithelial-mesenchymal transition (moesin (Msn)) was up-regulated in Hnf4a(H) mice. Moreover, we also found that miR-194/192 binds the 3-UTR of these mRNAs. siRNA knockdown of HNF4 suppressed miR-194/192 expression in human hepatocellular carcinoma (HCC) cells and resulted in up-regulation of their mRNA targets. Inhibition and overexpression experiments with miR-194/192 revealed that Gyg1, Setd5, Sumo2, Cln4b, and Rap2b are miR-194 targets, whereas Ereg, Alcam, and Msn are miR-192 targets. These findings reveal a novel HNF4 network controlled by miR-194/192 that may play a critical role in maintaining the hepatocyte-differentiated state by inhibiting expression of genes involved in dedifferentiation and tumorigenesis. These insights may contribute to the development of diagnostic markers for early HCC detection, and targeting of the miR-194/192 pathway could be useful for managing HCC.

    DOI: 10.1074/jbc.M117.785592

    Web of Science

    researchmap

  • Distant Bystander Effect of REIC/DKK3 Gene Therapy Through Immune System Stimulation in Thoracic Malignancies

    Ken Suzawa, Kazuhiko Shien, Huang Peng, Masakiyo Sakaguchi, Masami Watanabe, Shinsuke Hashida, Yuho Maki, Hiromasa Yamamoto, Shuta Tomida, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Hiromi Kumon, Shinichiro Miyoshi, Shinichi Toyooka

    ANTICANCER RESEARCH   37 ( 1 )   301 - 307   2017.1

     More details

    Language:English   Publisher:INT INST ANTICANCER RESEARCH  

    Background: Reduced expression in immortalized cell (REIC)/Dickkoph-3 (DKK3) is a tumor-suppressor gene, and its overexpression by adenovirus vector (Ad-REIC) exhibits a remarkable therapeutic effect on various human cancer types through a mechanism triggered by endoplasmic reticulum stress. Materials and Methods: We examined the direct anti-tumor effect of Ad-REIC gene therapy on lung cancer and malignant mesothelioma cell lines in vitro, and the distant bystander effect using immunocompetent mouse allograft models with bilateral flank tumors. Results: Ad-REIC treatment showed antitumor effect in many lung cancer and malignant mesothelioma cell lines in vitro. In an in vivo model, Ad-REIC treatment inhibited the growth not only of directly treated tumors but also of distant untreated tumors. By immunohistochemical analysis, infiltration of T-cells and natural killer (NK) cells and expression of the major histocompatibility complex (MHC) class I molecules were observed in bilateral tumors. Conclusion: Ad-REIC treatment not only had a direct antitumor effect but also an indirect bystander effect through stimulation of the immune system.

    DOI: 10.21873/anticanres.11321

    Web of Science

    researchmap

  • ß-1,3-galactosyl-O-glycosyl-glycoprotein ß-1,6-N-acetylglucosaminyltransferase 3 Increases MCAM Stability, Which Enhances S100A8/A9-Mediated Cancer Motility.

    Sumardika IW, Youyi C, Kondo E, Inoue Y, Ruma IMW, Murata H, Kinoshita R, Yamamoto KI, Tomida S, Shien K, Satoh H, Yamauchi A, Futami J, Putranto EW, Hibino T, Toyooka S, Nishibori M, Sakaguchi M

    Oncol Res   2017

     More details

  • 【アトピー性皮膚炎の病態研究】 Neuroplastin βとextracellular matrix metallo-protease inducerはS100A8/A9に対する機能的な受容体であり、ケラチノサイトの増殖とアトピーの皮膚炎症の増強に関与している

    阪口 政清, 山本, 真実, 宮井 雅史, 木下 理恵, 村田 等, 山本 健一, 森実 真, 岩月 啓氏, 許 南浩, 坪井 良治, 日比野 利彦

    臨床免疫・アレルギー科   67 ( 6 )   594 - 598   2017

     More details

  • S100-SPECT uncovers cellular and molecular events of pre-metastatic niche formation and following organ-specific cancer metastasis.

    Sakaguchi M

    Theranostics   7 ( 10 )   2649 - 2651   2017

     More details

  • Neuroplastinβとextracellular matrix metalloprotease inducerはS100A8/A9に対する機能的な受容体であり,ケラチノサイトの増殖とアトピーの皮膚炎症の増強に関与している. Critical role of novel receptors to S100A8/A9, EMMPRIN and NPTNβ, on keratinocyte growth and inflammation in atopic dermatitis.

    阪口政清, 山本真実, 宮井雅史, 松元 有羽子, 木下理恵, 村田 等, 山本健一, 森実 真, 岩月啓氏, 許 南浩, 坪井良治, 日比野 利彦

    臨床免疫   49 ( 6 )   594 - 598   2017

     More details

  • 転移先臓器を感知する受容体群の分子制御機構の解明とその応用. Advanced approach to prevent cancer metastasis based on S100A8/A9-mediated seed and soil theory Innovative approach to prevent cancer metastasis based on S100A8/A9-mediated seed and soil theory.

    阪口政清, 木下理恵, 村田 等, 山本健一, 許 南浩, 日比野 利彦

    月刊「細胞」6月号   49 ( 7 )   43 - 46   2017

     More details

  • 転移先臓器を感知する受容体群Innovative approach to prevent cancer metastasis based on S100A8/A9-mediated seed and soil theory.

    阪口政清, 木下理恵, 村田 等, 山本健一, 許 南浩, 日比野 利彦

    月刊「細胞」3月号   49 ( 3 )   39 - 42   2017

     More details

  • Active Secretion of Dimerized S100A11 Induced by the Peroxisome in Mesothelioma Cells. International journal

    Satomi Saho, Hiroki Satoh, Eisaku Kondo, Yusuke Inoue, Akira Yamauchi, Hitoshi Murata, Rie Kinoshita, Ken-Ichi Yamamoto, Junichiro Futami, Endy Widya Putranto, I Made Winarsa Ruma, I Wayan Sumardika, Chen Youyi, Ken Suzawa, Hiromasa Yamamoto, Junichi Soh, Shuta Tomida, Yoshihiko Sakaguchi, Ken Saito, Hidekazu Iioka, Nam-Ho Huh, Shinichi Toyooka, Masakiyo Sakaguchi

    Cancer microenvironment : official journal of the International Cancer Microenvironment Society   9 ( 2-3 )   93 - 105   2016.12

     More details

    Language:English  

    S100A11, a small Ca2+ binding protein, acts extracellularly as a mediator of cancer progression. That raises the question of how a protein that lacks the classical secretory signal is able to be secreted outside cells without being damaged. Some insights into this question have been obtained, and there has been accumulating evidence indicating a pivotal role of a non-classical vesicle-mediated pathway using lysosomes or peroxisomes for the protein secretion. To obtain a more precise insight into the secretory mechanism of S100A11, we first screened representative cancer cells exhibiting significantly active secretion of S100A11. From the results of profiling, we turned our attention to aggressive cancer mesothelioma cells. In mesothelioma cells, we found that abundant dimeric S100A11 was produced selectively in the peroxisome after transportation of monomeric S100A11 through an interaction with PEX14, a peroxisome membrane protein, resulting in peroxisomal secretion of dimerized S100A11. In an extracellular environment in vitro, dimerized S100A11 promoted mesothelial cell invasion indirectly with the help of fibroblast cells. Overall, the results indicate that the peroxisome functions as an essential vesicle for the production of dimerized S100A11 and the subsequent secretion of the protein from mesothelioma cells and that peroxisome-mediated secretion of dimerized S100A11 might play a critical role in mesothelioma progression in a tumor microenvironment.

    DOI: 10.1007/s12307-016-0185-2

    PubMed

    researchmap

  • Interaction of cytokeratin 19 head domain and HER2 in the cytoplasm leads to activation of HER2-Erk pathway

    Tomoaki Ohtsuka, Masakiyo Sakaguchi, Hiromasa Yamamoto, Shuta Tomida, Katsuyoshi Takata, Kazuhiko Shien, Shinsuke Hashida, Tomoko Miyata-Takata, Mototsugu Watanabe, Ken Suzawa, Junichi Soh, Chen Youyi, Hiroki Sato, Kei Namba, Hidejiro Torigoe, Kazunori Tsukuda, Tadashi Yoshino, Shinichiro Miyoshi, Shinichi Toyooka

    SCIENTIFIC REPORTS   6   39557 - 39557   2016.12

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    HER2 is a receptor tyrosine kinase and its upregulation via activating mutations or amplification has been identified in some malignant tumors, including lung cancers. Because HER2 can be a therapeutic target in HER2-driven malignancies, it is important to understand the molecular mechanisms of HER2 activation. In the current study, we identified that cytokeratin 19 (KRT19) binds to HER2 at the inside face of plasma membrane. HER2 and KRT19, which were concurrently introduced to a human embryonic kidney 293 T cells, revealed an association with each other and resulted in phosphorylation of HER2 with the subsequent activation of a downstream Erk-associated pathway. A binding assay revealed that both the NH2-terminal head domain of KRT19 and the COOH-terminal domain of HER2 were essential for their binding. To investigate the impact of the interaction between HER2 and KRT19 in lung cancer, we examined their expressions and localizations in lung cancers. We found that KRT19 was highly expressed in HER2-positive lung cancer cells, and KRT19 and HER2 were co-localized at the cell membrane. In conclusion, we found that KRT19 intracellularly binds to HER2, playing a critical role in HER2 activation.

    DOI: 10.1038/srep39557

    Web of Science

    researchmap

  • Identification of an S100A8 Receptor Neuroplastin-beta and its Heterodimer Formation with EMMPRIN

    Masakiyo Sakaguchi, Mami Yamamoto, Masashi Miyai, Tatsuo Maeda, Junichiro Hiruma, Hitoshi Murata, Rie Kinoshita, I. Made Winarsa Ruma, Endy Widya Putranto, Yusuke Inoue, Shin Morizane, Nam-Ho Huh, Ryoji Tsuboi, Toshihiko Hibino

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   136 ( 11 )   2240 - 2250   2016.11

     More details

    Language:English   Publisher:ELSEVIER SCIENCE INC  

    We previously reported a positive feedback loop between S100A8/A9 and proinflammatory cytokines mediated by extracellular matrix metalloproteinase inducer, an S100A9 receptor. Here, we identify neuroplastin-beta as an unreported S100A8 receptor. Neuroplastin-beta and extracellular matrix metalloproteinase inducer form homodimers and a heterodimer, and they are co-localized on the surface of cultured normal human keratinocytes. Knockdown of both receptors suppressed cell proliferation and proinflammatory cytokine induction. Upon stimulation with S100A8, neuroplastin-beta recruited GRB2 and activated extracellular signal-regulated kinase, resulting in keratinocyte proliferation. Keratinocyte proliferation in response to inflammatory stimuli was accelerated in involucrin promoter-driven S100A8 transgenic mice. Further, S100A8 and S100A9 were strongly up-regulated and co-localized in lesional skin of atopic dermatitis patients. Our results indicate that neuroplastin-beta and extracellular matrix metalloproteinase inducer form a functional heterodimeric receptor for S100A8/A9 heterodimer, followed by recruitment of specific adaptor molecules GRB2 and TRAF2, and this signaling pathway is involved in activation of both keratinocyte proliferation and skin inflammation in atopic skin. Suppression of this pathway might have potential for treatment of skin diseases associated with chronic inflammation such as atopic dermatitis.

    DOI: 10.1016/j.jid.2016.06.617

    Web of Science

    researchmap

  • RAGEはロイコトリエンB4第一受容体BLT1と機能的に相互作用する

    市木 貴子, 古賀 友紹, 奥野 利明, 佐伯 和子, 阪口 政清, 山本 靖彦, 横溝 岳彦

    日本生化学会大会プログラム・講演要旨集   89回   [2P - 089]   2016.9

     More details

    Language:Japanese   Publisher:(公社)日本生化学会  

    researchmap

  • A super gene expression system enhances the anti-glioma effects of adenovirus-mediated REIC/Dkk-3 gene therapy

    Tetsuo Oka, Kazuhiko Kurozumi, Yosuke Shimazu, Tomotsugu Ichikawa, Joji Ishida, Yoshihiro Otani, Toshihiko Shimizu, Yusuke Tomita, Masakiyo Sakaguchi, Masami Watanabe, Yasutomo Nasu, Hiromi Kumon, Isao Date

    SCIENTIFIC REPORTS   6   33319 - 33319   2016.9

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    Reduced expression in immortalized cells/Dickkopf-3 (REIC/Dkk-3) is a tumor suppressor and therapeutic gene in many human cancers. Recently, an adenovirus REIC vector with the super gene expression system (Ad-SGE-REIC) was developed to increase REIC/Dkk-3 expression and enhance therapeutic effects compared with the conventional adenoviral vector (Ad-CAG-REIC). In this study, we investigated the in vitro and in vivo effects of Ad-SGE-REIC on malignant glioma. In U87 Delta EGFR and GL261 glioma cells, western blotting confirmed that robust upregulation of REIC/Dkk-3 expression occurred in Ad-SGE-REIC-transduced cells, most notably after transduction at a multiplicity of infection of 10. Cytotoxicity assays showed that Ad-SGE-REIC resulted in a time-dependent and significant reduction in the number of malignant glioma cells attaching to the bottom of culture wells. Xenograft and syngeneic mouse intracranial glioma models treated with Ad-SGE-REIC had significantly longer survival than those treated with the control vector Ad-LacZ or with Ad-CAG-REIC. This study demonstrated the anti-glioma effect of Ad-SGE-REIC, which may represent a promising strategy for the treatment of malignant glioma.

    DOI: 10.1038/srep33319

    Web of Science

    researchmap

  • Novel REIC/Dkk-3-encoding adenoviral vector as a promising therapeutic agent for pancreatic cancer

    H. Sawahara, H. Shiraha, D. Uchida, H. Kato, T. Nagahara, M. Iwamuro, J. Kataoka, S. Horiguchi, M. Watanabe, M. Sakaguchi, A. Takaki, K. Nouso, Y. Nasu, H. Kumon, H. Okada

    CANCER GENE THERAPY   23 ( 8 )   278 - 283   2016.8

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    Reduced expression in immortalized cells (REIC)/dickkopf-3 (Dkk-3), a tumor suppressor gene, is downregulated in various cancers. We previously reported the tumor-inhibitory effects of the REIC/Dkk-3 gene, delivered by a conventional adenoviral vector (Ad-CAG-REIC) in pancreatic cancer. Here, we developed an Ad-REIC vector with a novel gene expression system, termed the super gene expression (SGE) system, and assessed its therapeutic effects relative to those of Ad-CAG-REIC in pantreatic cancer cells. Human pancreatic cancer cell lines ASPC1 and MIAPaCa2 were used. REIC/Dkk-3 expression was assessed by western blot analysis. Relative cell viability and apoptotic effects were examined in vitro. The anti-tumor effects of Ad-REIC treatment were assessed in the mouse xenograft model. Compared with Ad-CAG-REIC, Ad-SGE-REIC elicited a significant increase in REIC protein expression in the cells studied, Relative to Ad-CAG-REIC, Ad-SGE-REIC reduced cell viability and induced apoptosis in the ASPC1 and MIAPaCa2 cell lines in vitro, and achieved superior tumor growth inhibition in the mouse xenograft model. Compared with conventional Ad-REIC agents, Ad-SGE-REIC provided enhanced inhibitory effects against tumor growth. Our results indicate that Ad-SGE-REIC is an innovative therapeutic tool for pancreatic cancer.

    DOI: 10.1038/cgt.2016.31

    Web of Science

    researchmap

  • MCAM, as a novel receptor for S100A8/A9, mediates progression of malignant melanoma through prominent activation of NF-κB and ROS formation upon ligand binding. International journal

    I Made Winarsa Ruma, Endy Widya Putranto, Eisaku Kondo, Hitoshi Murata, Masami Watanabe, Peng Huang, Rie Kinoshita, Junichiro Futami, Yusuke Inoue, Akira Yamauchi, I Wayan Sumardika, Chen Youyi, Ken-Ichi Yamamoto, Yasutomo Nasu, Masahiro Nishibori, Toshihiko Hibino, Masakiyo Sakaguchi

    Clinical & experimental metastasis   33 ( 6 )   609 - 27   2016.8

     More details

    Language:English   Publisher:SPRINGER  

    The dynamic interaction between tumor cells and their microenvironment induces a proinflammatory milieu that drives cancer development and progression. The S100A8/A9 complex has been implicated in chronic inflammation, tumor development, and progression. The cancer microenvironment contributes to the up-regulation of this protein complex in many invasive tumors, which is associated with the formation of pre-metastatic niches and poor prognosis. Changing adhesive preference of cancer cells is at the core of the metastatic process that governs the reciprocal interactions of cancer cells with the extracellular matrices and neighboring stromal cells. Cell adhesion molecules (CAMs) have been confirmed to have high-level expression in various highly invasive tumors. The expression and function of CAMs are profoundly influenced by the extracellular milieu. S100A8/A9 mediates its effects by binding to cell surface receptors, such as heparan sulfate, TLR4 and RAGE on immune and tumor cells. RAGE has recently been identified as an adhesion molecule and has considerably high identity and similarity to ALCAM and MCAM, which are frequently over-expressed on metastatic malignant melanoma cells. In this study, we demonstrated that ALCAM and MCAM also function as S100A8/A9 receptors as does RAGE and induce malignant melanoma progression by NF-κB activation and ROS formation. Notably, MCAM not only activated NF-κB more prominently than ALCAM and RAGE did but also mediated intracellular signaling for the formation of lung metastasis. MCAM is known to be involved in malignant melanoma development and progression through several mechanisms. Therefore, MCAM is a potential effective target in malignant melanoma treatment.

    DOI: 10.1007/s10585-016-9801-2

    Web of Science

    PubMed

    researchmap

  • An efficient method for the preparation of preferentially heterodimerized recombinant S100A8/A9 coexpressed in Escherichia coli

    Junichiro Futami, Yuki Atago, Akari Azuma, Endy Widya Putranto, Rie Kinoshita, Hitoshi Murata, Masakiyo Sakaguchi

    Biochemistry and Biophysics Reports   6   94 - 100   2016.7

     More details

    Language:English   Publisher:Elsevier B.V.  

    It is now known that multicomponent protein assemblies strictly regulate many protein functions. The S100 protein family is known to play various physiological roles, which are associated with alternative complex formations. To prepare sufficient amounts of heterodimeric S100A8 and S100A9 proteins, we developed a method for bicistronic coexpression from a single-vector system using Escherichia coli cells as a host. The complex formation between S100A8 and S100A9 appears to be dependent on the thermodynamic stability of the protein during expression. The stable S100A8/A9 heterodimer complex spontaneously formed during coexpression, and biologically active samples were purified by cation-exchange chromatography. Semi-stable homodimers of S100A8 and S100A9 were also formed when expressed individually. These results suggest that the assembly of S100 protein complexes might be regulated by expression levels of partner proteins in vivo. Because protein assembly occurs rapidly after protein synthesis, coexpression of relevant proteins is crucial for the design of multicomponent recombinant protein expression systems.

    DOI: 10.1016/j.bbrep.2016.03.009

    Scopus

    researchmap

  • Histidine-Rich Glycoprotein Prevents Septic Lethality through Regulation of Immunothrombosis and Inflammation

    Hidenori Wake, Shuji Mori, Keyue Liu, Yuta Morioka, Kiyoshi Teshigawara, Masakiyo Sakaguchi, Kosuke Kuroda, Yuan Gao, Hideo Takahashi, Aiji Ohtsuka, Tadashi Yoshino, Hiroshi Morimatsu, Masahiro Nishibori

    EBIOMEDICINE   9   180 - 194   2016.7

     More details

    Language:English   Publisher:ELSEVIER SCIENCE BV  

    Sepsis is a major cause of death worldwide. We show that a plasma protein histidine-rich glycoprotein (HRG) was decreased significantly in septic mice with cecal ligation and puncture (CLP) and supplementary treatment of septic mice with exogenous HRG improved survival, with strong inhibition of tight attachment of neutrophils to pulmonary vasculatures, subsequent immunothrombosis, DIC state, lung inflammation, hypercytokinemia, and activation of vascular endothelial cells (VECs). In contrast, knockdown of HRG by siRNA exacerbated lethality. Purified human HRG reversibly induced morphological changes in human neutrophils in vitro; induction of spherical shape with reduced microvilli and adhesiveness to VECs. HRG maintained the passage of neutrophils through microcapillaries and abolished production of reactive oxygen species. These results suggested that the supplementary therapy with HRG may provide a novel strategy for the treatment of sepsis through suppression of excessive systemic inflammation and immunothrombosis by keeping circulating neutrophils quiescent and preventing uncontrolled activation of VECs. (C) 2016 The Authors. Published by Elsevier B.V.

    DOI: 10.1016/j.ebiom.2016.06.003

    Web of Science

    researchmap

  • Antitumor effect of afatinib, as a human epidermal growth factor receptor 2-targeted therapy, in lung cancers harboring HER2 oncogene alterations

    Ken Suzawa, Shinichi Toyooka, Masakiyo Sakaguchi, Mizuki Morita, Hiromasa Yamamoto, Shuta Tomida, Tomoaki Ohtsuka, Mototsugu Watanabe, Shinsuke Hashida, Yuho Maki, Junichi Soh, Hiroaki Asano, Kazunori Tsukuda, Shinichiro Miyoshi

    CANCER SCIENCE   107 ( 1 )   45 - 52   2016.1

     More details

    Language:English   Publisher:WILEY  

    Human epidermal growth factor receptor 2 (HER2) is a member of the HER family of proteins containing four receptor tyrosine kinases. It plays an important role in the pathogenesis of certain human cancers. In non-small-cell lung cancer (NSCLC), HER2 amplification or mutations have been reported. However, little is known about the benefit of HER2-targeted therapy for NSCLCs harboring HER2 alterations. In this study, we investigated the antitumor effect of afatinib, an irreversible epidermal growth factor receptor (EGFR)-HER2 dual inhibitor, in lung cancers harboring HER2 oncogene alterations, including novel HER2 mutations in the transmembrane domain, which we recently identified. Normal bronchial epithelial cells, BEAS-2B, ectopically overexpressing wild-type HER2 or mutants (A775insYVMA, G776VC, G776LC, P780insGSP, V659E, and G660D) showed constitutive autophosphorylation of HER2 and activation of downstream signaling. They were sensitive to afatinib, but insensitive to gefitinib. Furthermore, we examined the antitumor activity of afatinib and gefitinib in several NSCLC cell lines, and investigated the association between their genetic alterations and sensitivity to afatinib treatment. In HER2-altered NSCLC cells (H2170, Calu-3, and H1781), afatinib downregulated the phosphorylation of HER2 and EGFR as well as their downstream signaling, and induced an antiproliferative effect through G(1) arrest and apoptotic cell death. In contrast, HER2- or EGFR-non-dependent NSCLC cells were insensitive to afatinib. In addition, these effects were confirmed in vivo by using a xenograft mouse model of HER2-altered lung cancer cells. Our results suggest that afatinib is a therapeutic option as a HER2-targeted therapy for NSCLC harboring HER2 amplification or mutations.

    DOI: 10.1111/cas.12845

    Web of Science

    researchmap

  • Modulation of leukotriene B4 receptor 1 signaling by receptor for advanced glycation end products (RAGE).

    Ichiki T, Koga T, Okuno T, Saeki K, Yamamoto Y, Yamamoto H, Sakaguchi M, Yokomizo T

    FASEB J   30 ( 5 )   1811 - 1822   2016

     More details

  • Hepatocyte Nuclear Factor 4 alpha Controls Iron Metabolism and Regulates Transferrin Receptor 2 in Mouse Liver

    Shunsuke Matsuo, Masayuki Ogawa, Martina U. Muckenthaler, Yumiko Mizui, Shota Sasaki, Takafumi Fujimura, Masayuki Takizawa, Nagayuki Ariga, Hiroaki Ozaki, Masakiyo Sakaguchi, Frank J. Gonzalez, Yusuke Inoue

    JOURNAL OF BIOLOGICAL CHEMISTRY   290 ( 52 )   30855 - 30865   2015.12

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Iron is an essential element in biological systems, but excess iron promotes the formation of reactive oxygen species, resulting in cellular toxicity. Several iron-related genes are highly expressed in the liver, a tissue in which hepatocyte nuclear factor 4 alpha (HNF4 alpha) plays a critical role in controlling gene expression. Therefore, the role of hepatic HNF4 alpha in iron homeostasis was examined using liver-specific HNF4 alpha-null mice (Hnf4a(Delta H) mice). Hnf4a(Delta H) mice exhibit hypoferremia and a significant change in hepatic gene expression. Notably, the expression of transferrin receptor 2 (Tfr2) mRNA was markedly decreased in Hnf4a(Delta H) mice. Promoter analysis of the Tfr2 gene showed that the basal promoter was located at a GC-rich region upstream of the transcription start site, a region that can be transactivated in an HNF4 alpha-independent manner. HNF4 alpha-dependent expression of Tfr2 was mediated by a proximal promoter containing two HNF4 alpha-binding sites located between the transcription start site and the translation start site. Both the GC-rich region of the basal promoter and the HNF4 alpha-binding sites were required for maximal transactivation. Moreover, siRNA knockdown of HNF4 alpha suppressed TFR2 expression in human HCC cells. These results suggest that Tfr2 is a novel target gene for HNF4 alpha, and hepatic HNF4 alpha plays a critical role in iron homeostasis.

    DOI: 10.1074/jbc.M115.694414

    Web of Science

    researchmap

  • NRF2 Regulates PINK1 Expression under Oxidative Stress Conditions

    Hitoshi Murata, Hitoshi Takamatsu, Sulai Lie, Ken Kataoka, Nam-ho Huh, Masakiyo Sakaguchi

    PLoS One   10 ( 11 )   e0142438-e0142438   2015.11

     More details

    Language:English   Publisher:PUBLIC LIBRARY SCIENCE  

    Mutations of the PTEN-induced putative kinase 1 (PINK1) gene are a cause of autosomal recessive forms of Parkinson's disease. Recent studies have revealed that PINK1 is an essential factor for controlling mitochondrial quality, and that it protects cells from oxidative stresses. Although there has been considerable progress in the elucidation of various aspects of PINK1 protein regulation such as activation, stability and degradation, the transcriptional regulation of PINK1 mRNA under stress conditions remains unclear. In this study, we found that nuclear factor (erythroid-derived 2)-like 2 (NRF2), an antioxidant transcription factor, regulates PINK1 expression under oxidative stress conditions. Damaged mitochondria arising from stress conditions induced NRF2-dependent transcription of the PINK1 gene through production of reactive oxygen species (ROS). Either an ROS scavenger or forced expression of KEAP1, a potent inhibitory partner to NRF2, restricted PINK1 expression induced by activated NRF2. Transcriptionally up-regulated PINK1 diminished oxidative stress-associated cell death. The results indicate that PINK1 expression is positively regulated by NRF2 and that the NRF2-PINK1 signaling axis is deeply involved in cell survival.

    DOI: 10.1371/journal.pone.0142438

    Web of Science

    researchmap

  • TAE226, a Bis-Anilino Pyrimidine Compound, Inhibits the EGFR-Mutant Kinase Including T790M Mutant to Show Anti-Tumor Effect on EGFR-Mutant Non-Small Cell Lung Cancer Cells

    Hiroki Otani, Hiromasa Yamamoto, Munenori Takaoka, Masakiyo Sakaguchi, Junichi Soh, Masaru Jida, Tsuyoshi Ueno, Takafumi Kubo, Hiroaki Asano, Kazunori Tsukuda, Katsuyuki Kiura, Shinji Hatakeyama, Eiji Kawahara, Yoshio Naomoto, Shinichiro Miyoshi, Shinichi Toyooka

    PLOS ONE   10 ( 6 )   e0129838-e0129838   2015.6

     More details

    Language:English   Publisher:PUBLIC LIBRARY SCIENCE  

    TAE226, a bis-anilino pyrimidine compound, has been developed as an inhibitor of focal adhesion kinase (FAK) and insulin-like growth factor-I receptor (IGF-IR). In this study, we investigated the effect of TAE226 on non-small-cell lung cancer (NSCLC), especially focusing on the EGFR mutational status. TAE226 was more effective against cells with mutant EGFR, including the T790M mutant, than against cells with wild-type one. TAE226 preferentially inhibited phospho-EGFR and its downstream signaling mediators in the cells with mutant EGFR than in those with wild-type one. Phosphorylation of FAK and IGF-IR was not inhibited at the concentration at which the proliferation of EGFR-mutant cells was inhibited. Results of the in vitro binding assay indicated significant differences in the affinity for TAE226 between the wild-type and L858R (or delE746_A750) mutant, and the reduced affinity of ATP to the L858R (or delE746_A750) mutant resulted in good responsiveness of the L858R (or delE746_A750) mutant cells to TAE226. Of interest, the L858R/T790M or delE746_A750/T790M mutant enhanced the binding affinity for TAE226 compared with the L858R or delE746_A750 mutant, resulting in the effectiveness of TAE226 against T790M mutant cells despite the T790M mutation restoring the ATP affinity for the mutant EGFR close to that for the wild-type. TAE226 also showed higher affinity of about 15-fold for the L858R/T790M mutant than for the wild-type one by kinetic interaction analysis. The antitumor effect against EGFR-mutant tumors including T790M mutation was confirmed in mouse models without any significant toxicity. In summary, we showed that TAE226 inhibited the activation of mutant EGFR and exhibited anti-proliferative activity against NSCLCs carrying EGFR mutations, including T790M mutation.

    DOI: 10.1371/journal.pone.0129838

    Web of Science

    researchmap

  • The cysteine-rich core domain of REIC/Dkk-3 is critical for its effect on monocyte differentiation and tumor regression

    Rie Kinoshita, Masami Watanabe, Peng Huang, Shun-Ai Li, Masakiyo Sakaguchi, Hiromi Kumon, Junichiro Futami

    ONCOLOGY REPORTS   33 ( 6 )   2908 - 2914   2015.6

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Reduced expression in immortalized cells (REIC)/Dickkopf (Dkk)-3 is a tumor-suppressor gene and has been studied as a promising therapeutic gene for cancer gene therapy. Intratumoral injection of an adenovirus vector carrying the human REIC/Dkk-3 gene (Ad-REIC) elicits cancer cell-specific apoptosis and anticancer immune responses. The cytokine-like effect of secretory REIC/Dkk-3 on the induction of dendritic cell (DC)-like cell differentiation from monocytes plays a role in systemic anticancer immunity. In the present study, we generated recombinant full-length and N-terminally truncated REIC/Dkk-3 to characterize the biological activity of the protein. During the purification procedure, we identified a 17 kDa cysteine-rich stable product (C17-REIC) showing limited degradation. Further analysis showed that the C17-REIC domain was sufficient for the induction of DC-like cell differentiation from monocytes. Concomitant with the differentiation of DCs, the REIC/Dkk-3 protein induced the phosphorylation of glycogen synthase kinase 3 beta (GSK-3 beta) and signal transducers and activators of transcription (STAT) at a level comparable to that of granulocyte/macrophage colony-stimulating factor. In a mouse model of subcutaneous renal adenocarcinoma, intraperitoneal injection of full-length and C17-REIC proteins exerted anticancer effects in parallel with the activation of immunocompetent cells such as DCs and cytotoxic T lymphocytes in peripheral blood. Taken together, our results indicate that the stable cysteine-rich core region of REIC/Dkk-3 is responsible for the induction of anticancer immune responses. Because REIC/Dkk-3 is a naturally circulating serum protein, the upregulation REIC/Dkk-3 protein expression could be a promising option for cancer therapy.

    DOI: 10.3892/or.2015.3885

    Web of Science

    researchmap

  • Antitumor effect of afatinib, as a HER2-targeted therapy, in lung cancers harboring HER2 oncogene alterations.

    Suzawa K, Toyooka S, Sakaguchi M, Morita M, Yamamoto H, Tomida S, Ohtsuka T, Watanabe M, Hashida S, Maki Y, Soh J, Asano H, Tsukuda K, Miyoshi S

    Cancer Sci   cas.12845-cas.12845   2015

     More details

  • Potential role of tumor suppressor gene REIC/Dkk-3 as a regulator of skin tissue inflammation

    Y. Ayabe, T. Tsuruda, M. Sakaguchi, K. Kataoka

    MOLECULAR BIOLOGY OF THE CELL   26   2015

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • 肺腺癌におけるHER2膜貫通領域の遺伝子変異と機能解析

    山本寛斉, 阪口政清, 諏澤憲, 大塚智昭, 枝園和彦, 橋田真輔, 渡邉元嗣, 牧佑歩, 宗淳一, 岡田真典, 伊賀徳周, 三好健太郎, 杉本誠一郎, 山根正修, 大藤剛宏, 三好新一郎, 豊岡伸一

    日本肺癌学会総会号   55th ( 5 )   362 - 362   2014.10

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • Anti-cancer effects of REIC/Dkk-3-encoding adenoviral vector for the treatment of non-small cell lung cancer

    Ken Suzawa, Shinichi Toyooka, Kazuhiko Shien, Norimitsu Tanaka, Masami Watanabe, Junichi Soh, Masakiyo Sakaguchi, Keitaro Matsuo, Hiromasa Yamamoto, Masashi Furukawa, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Nam-Ho Huh, Shinichiro Miyoshi, Hiromi Kumon

    CANCER RESEARCH   74 ( 19 )   2014.10

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER ASSOC CANCER RESEARCH  

    DOI: 10.1158/1538-7445.AM2014-2879

    Web of Science

    researchmap

  • DNAX-activating Protein 10 (DAP10) Membrane Adaptor Associates with Receptor for Advanced Glycation End Products (RAGE) and Modulates the RAGE-triggered Signaling Pathway in Human Keratinocytes

    Masakiyo Sakaguchi, Hitoshi Murata, Yumi Aoyama, Toshihiko Hibino, Endy Widya Putranto, I. Made Winarsa Ruma, Yusuke Inoue, Yoshihiko Sakaguchi, Ken-ichi Yamamoto, Rie Kinoshita, Junichiro Futami, Ken Kataoka, Keiji Iwatsuki, Nam-ho Huh

    JOURNAL OF BIOLOGICAL CHEMISTRY   289 ( 34 )   23389 - 23402   2014.8

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Background: RAGE receptor plays a critical role in many inflammatory disorders. Results: Functional interaction between RAGE and DAP10 coordinately regulates S100A8/A9-mediated cell survival. Conclusion: DAP10 membrane adaptor is critically involved in RAGE-mediated survival signaling upon S100A8/A9 binding. Significance: This is the first report demonstrating that RAGE-mediated survival signaling is critically regulated by DAP10 interaction.
    The receptor for advanced glycation end products (RAGE) is involved in the pathogenesis of many inflammatory, degenerative, and hyperproliferative diseases, including cancer. Previously, we revealed mechanisms of downstream signaling from ligand-activated RAGE, which recruits TIRAP/MyD88. Here, we showed that DNAX-activating protein 10 (DAP10), a transmembrane adaptor protein, also binds to RAGE. By artificial oligomerization of RAGE alone or RAGE-DAP10, we found that RAGE-DAP10 heterodimer formation resulted in a marked enhancement of Akt activation, whereas homomultimeric interaction of RAGE led to activation of caspase 8. Normal human epidermal keratinocytes exposed to S100A8/A9, a ligand for RAGE, at a nanomolar concentration mimicked the pro-survival response of RAGE-DAP10 interaction, although at a micromolar concentration, the cells mimicked the pro-apoptotic response of RAGE-RAGE. In transformed epithelial cell lines, A431 and HaCaT, in which endogenous DAP10 was overexpressed, and S100A8/A9, even at a micromolar concentration, led to cell growth and survival due to RAGE-DAP10 interaction. Functional blocking of DAP10 in the cell lines abrogated the Akt phosphorylation from S100A8/A9-activated RAGE, eventually leading to an increase in apoptosis. Finally, S100A8/A9, RAGE, and DAP10 were overexpressed in the psoriatic epidermis. Our findings indicate that the functional interaction between RAGE and DAP10 coordinately regulates S100A8/A9-mediated survival and/or apoptotic response of keratinocytes.

    DOI: 10.1074/jbc.M114.573071

    Web of Science

    researchmap

  • Extract of Cordyceps militaris inhibits angiogenesis and suppresses tumor growth of human malignant melanoma cells

    I. Made Winarsa Ruma, Endy Widya Putranto, Eisaku Kondo, Risayo Watanabe, Ken Saito, Yusuke Inoue, Ken-Ichi Yamamoto, Susumu Nakata, Masaji Kaihata, Hitoshi Murata, Masakiyo Sakaguchi

    INTERNATIONAL JOURNAL OF ONCOLOGY   45 ( 1 )   209 - 218   2014.7

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Angiogenesis is essential for tumor development and metastasis. Among several angiogenic factors, vascular endothelial growth factor receptor (VEGF) is important for tumor-derived angiogenesis and commonly overexpressed in solid tumors. Thus, many antitumor strategies targeting VEGF have been developed to inhibit cancer angiogenesis, offering insights into the successful treatment of solid cancers. However, there are a number of issues such as harmful effects on normal vascularity in clinical trials. Taking this into consideration, we employed Cordyceps militaris as an antitumor approach due to its biological safety in vivo. The herbal medicinal mushroom Cordyceps militaris has been reported to show potential anticancer properties including anti-angiogenic capacity; however, its concrete properties have yet to be fully demonstrated. In this study, we aimed to elucidate the biological role of Cordyceps militaris extract in tumor cells, especially in regulating angiogenesis and tumor growth of a human malignant melanoma cell line. We demonstrated that Cordyceps militaris extract remarkably suppressed tumor growth via induction of apoptotic cell death in culture that links to the abrogation of VEGF production in melanoma cells. This was followed by mitigation of Akt1 and GSK-3 beta activation, while p38 alpha phosphorylation levels were increased. Extract treatment in mouse model xenografted with human melanoma cells resulted in a dramatic antitumor effect with downregulation of VEGF expression. The results suggest that suppression of tumor growth by Cordyceps militaris extract is, at least, mediated by its anti-angiogenicity and apoptosis induction capacities. Cordyceps militaris extract may be a potent antitumor herbal drug for solid tumors.

    DOI: 10.3892/ijo.2014.2397

    Web of Science

    researchmap

  • Dramatic Increase in Expression of a Transgene by Insertion of Promoters Downstream of the Cargo Gene

    Masakiyo Sakaguchi, Masami Watanabe, Rie Kinoshita, Haruki Kaku, Hideo Ueki, Junichiro Futami, Hitoshi Murata, Yusuke Inoue, Shun-Ai Li, Peng Huang, Endy Widya Putranto, I. Made Winarsa Ruma, Yasutomo Nasu, Hiromi Kumon, Nam-ho Huh

    MOLECULAR BIOTECHNOLOGY   56 ( 7 )   621 - 630   2014.7

     More details

    Language:English   Publisher:HUMANA PRESS INC  

    For expression of genes in mammalian cells, various vectors have been developed using promoters including CMV, EF-1 alpha, and CAG promoters and have been widely used. However, such expression vectors sometimes fail to attain sufficient expression levels depending on the nature of cargo genes and/or on host cell types. In the present study, we aimed to develop a potent promoter system that enables high expression levels of cargo genes ubiquitously in many different cell types. We found that insertion of an additional promoter downstream of a cargo gene greatly enhanced the expression levels. Among the constructs we tested, C-TSC cassette (C: CMV-RU5' located upstream; TSC: another promoter unit composed of triple tandem promoters, hTERT, SV40, and CMV, located downstream of the cDNA plus a polyadenylation signal) had the most potent capability, showing far higher efficiency than that of potent conventional vector systems. The results indicate that the new expression system is useful for production of recombinant proteins in mammalian cells and for application as a gene therapeutic measure.

    DOI: 10.1007/s12033-014-9738-0

    Web of Science

    researchmap

  • S100A11 is required for efficient plasma membrane repair and survival of invasive cancer cells

    Jyoti K. Jaiswal, Stine P. Lauritzen, Luana Scheffer, Masakiyo Sakaguchi, Jakob Bunkenborg, Sanford M. Simon, Tuula Kallunki, Marja Jaattela, Jesper Nylandsted

    NATURE COMMUNICATIONS   5   a3795-a3795   2014.5

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    Cell migration and invasion require increased plasma membrane dynamics and ability to navigate through dense stroma, thereby exposing plasma membrane to tremendous physical stress. Yet, it is largely unknown how metastatic cancer cells acquire an ability to cope with such stress. Here we show that S100A11, a calcium-binding protein upregulated in a variety of metastatic cancers, is essential for efficient plasma membrane repair and survival of highly motile cancer cells. Plasma membrane injury-induced entry of calcium into the cell triggers recruitment of S100A11 and Annexin A2 to the site of injury. We show that S100A11 in a complex with Annexin A2 helps reseal the plasma membrane by facilitating polymerization of cortical F-actin and excision of the damaged part of the plasma membrane. These data reveal plasma membrane repair in general and S100A11 and Annexin A2 in particular as new targets for the therapy of metastatic cancers.

    DOI: 10.1038/ncomms4795

    Web of Science

    researchmap

  • 超高効率遺伝子発現システムを用いた次世代REIC遺伝子治療の開発

    渡部 昌実, 阪口 政清, 賀来 春紀, 佐々木 克己, 高本 篤, 有吉 勇一, 杉本 盛人, 江原 伸, 渡辺 豊彦, 許 南浩, 那須 保友, 公文 裕巳

    日本泌尿器科学会総会   102回   621 - 621   2014.4

     More details

    Language:Japanese   Publisher:(一社)日本泌尿器科学会総会事務局  

    researchmap

  • A novel gene expression system strongly enhances the anticancer effects of a REIC/Dkk-3-encoding adenoviral vector

    Masami Watanabe, Masakiyo Sakaguchi, Rie Kinoshita, Haruki Kaku, Yuichi Ariyoshi, Hideo Ueki, Ryuta Tanimoto, Shin Ebara, Kazuhiko Ochiai, Junichiro Futami, Shun-Ai Li, Peng Huang, Yasutomo Nasu, Nam-Ho Huh, Hiromi Kumon

    ONCOLOGY REPORTS   31 ( 3 )   1089 - 1095   2014.3

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Gene expression systems with various promoters, including the cytomegalovirus (CMV) promoter, have been developed to increase the gene expression in a variety of normal and cancer cells. In particular, in the clinical trials of cancer gene therapy, a more efficient and robust gene expression system is required to achieve sufficient therapeutic outcomes. By inserting the triple translational enhancer sequences of human telomerase reverse transcriptase (hTERT), Simian virus 40 (SV40) and CMV downstream of the sequence of the BGH polyA, we were able to develop a novel gene expression system that significantly enhances the expression of the genes of interest. We termed this novel gene expression cassette the super gene expression (SGE) system, and herein verify the utility of the SGE cassette for a replication-deficient adenoviral vector. We newly developed an adenoviral vector expressing the tumor suppressor, reduced expression in immortalized cells (REIC)/Dickkopf-3 (Dkk-3), based on the CMV promoter-driven SGE system (Ad-SGE-REIC) and compared the therapeutic utility of Ad-SGE-REIC with that of the conventional adenoviral vectors (Ad-CMV-REIC or Ad-CAG-REIC). The results demonstrated that the CMV promoter-SGE system allows for more potent gene expression, and that the Ad-SGE-REIC is superior to conventional adenoviral systems in terms of the REIC protein expression and therapeutic effects. Since the SGE cassette can be applied for the expression of various therapeutic genes using various vector systems, we believe that this novel system will become an innovative tool in the field of gene expression and gene therapy.

    DOI: 10.3892/or.2013.2958

    Web of Science

    researchmap

  • Coxsackie and adenovirus receptor is a critical regulator for the survival and growth of oral squamous carcinoma cells

    K. Saito, M. Sakaguchi, H. Iioka, M. Matsui, H. Nakanishi, N. H. Huh, E. Kondo

    ONCOGENE   33 ( 10 )   1274 - 1286   2014.3

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    Coxsackie and adenovirus receptor (CAR) is essential for adenovirus infection to target cells, and its constitutive expression in various cancerous and normal tissues has been reported. Recently, the biological role of CAR in human cancers of several different origins has been investigated with respect to tumor progression, metastasis and tumorigenesis. However, its biological function in tumor cells remains controversial. Here we report the critical role of CAR in growth regulation of oral squamous cell carcinomas (SCCs) in vitro and in vivo via the specific interaction with Rho-associated protein kinase (ROCK). Loss of endogenous CAR expression by knockdown using specific small interfering RNA (siRNA) against CAR facilitates growth suppression of SCC cells due to cell dissociation, followed by apoptosis. The consequent morphological reaction was reminiscent of anoikis, rather than epithelial-mesenchymal transition, and the dissociation of oral SCC cells was triggered not by lack of contact with extracellular matrix, but by loss of cell-to-cell contact caused by abnormal translocation of E-cadherin from surface membrane to cytoplasm. Immunoprecipitation assays of the CAR-transfected oral SCC cell line, HSC-2, with or without ROCK inhibitor (Y-27632) revealed that CAR directly associates with ROCK! and ROCKII, which results in inhibition of ROCK activity and contributes to maintenance of cell-to-cell adhesion for their growth and survival. Based on these findings, in vivo behavior of CAR-downregulated HSC-2 cells from siRNA knockdown was compared with that of normally CAR-expressing cells in intraperitoneally xenografted mouse models. The mice engrafted with CAR siRNA-pretreated HSC-2 cells showed poor formation of metastatic foci in contrast to those implanted with the control siRNA-pretreated cells. Thus, CAR substantially has an impact on growth and survival of oral SCC cells as a negative regulator of ROCK in vitro and in vivo.

    DOI: 10.1038/onc.2013.66

    Web of Science

    researchmap

  • Anti-Cancer Effects of REIC/Dkk-3-encoding Adenoviral Vector for the Treatment of Non-small Cell Lung Cancer

    Kazuhiko Shien, Norimitsu Tanaka, Masami Watanabe, Junichi Soh, Masakiyo Sakaguchi, Keitaro Matsuo, Hiromasa Yamamoto, Masashi Furukawa, Hiroaki Asano, Kazunori Tsukuda, Yasutomo Nasu, Nam-Ho Huh, Shinichiro Miyoshi, Hiromi Kumon, Shinichi Toyooka

    PLOS ONE   9 ( 2 )   e87900-e87900   2014.2

     More details

    Language:English   Publisher:PUBLIC LIBRARY SCIENCE  

    Objectives: REIC/Dkk-3 is down-regulated in a broad range of human cancer cells and is considered to function as a tumor suppressor. We previously reported that REIC/Dkk-3-expressing adenovirus vector (Ad-REIC) induced endoplasmic reticulum (ER) stress and cancer-specific apoptosis in human prostate cancer. In this study, we examined the therapeutic impact of Ad-REIC on non-small cell lung cancer (NSCLC).
    Materials and Methods: We examined the anti-tumor effect of Ad-REIC on 25 NSCLC cell lines in vitro and A549 cells in vivo. Two of these cell lines were artificially established as EGFR-tyrosine kinase inhibitor (TKI) resistant sublines.
    Results: Ad-REIC-treatment inhibited the cell viability by 40% or more in 13 (52%) of the 25 cell lines at multiplicity of infection (MOI) of 20 (20 MOI). These cell lines were regarded as being highly sensitive cells. The cell viability of a nonmalignant immortalized cell line, OUMS-24, was not inhibited at 200 MOI of Ad-REIC. The effects of Ad-REIC on EGFR-TKI resistant sublines were equivalent to those in the parental cell lines. Here, we demonstrated that Ad-REIC treatment activated c-Jun N-terminal kinase (JNK) in NSCLC cell lines, indicating the induction of ER stress with GRP78/BiP (GRP78) up-regulation and resulting in apoptosis. A single intratumoral injection of Ad-REIC significantly inhibited the tumorigenic growth of A549 cells in vivo. As predictive factors of sensitivity for Ad-REIC treatment in NSCLC, we examined the expression status of GRP78 and coxsackievirus and adenovirus receptor (CAR). We found that the combination of the GRP78 and CAR expressional statuses may be used as a predictive factor for Ad-REIC sensitivity in NSCLC cells.
    Conclusion: Ad-REIC induced JNK activation and subsequent apoptosis in NSCLC cells. Our study indicated that Ad-REIC has therapeutic potential against NSCLC and that the expression statuses of GRP78 and CAR may predict a potential therapeutic benefit of Ad-REIC.

    DOI: 10.1371/journal.pone.0087900

    Web of Science

    researchmap

  • Preclinical Evaluation of MicroRNA-34b/c Delivery for Malignant Pleural Mesothelioma

    Tsuyoshi Ueno, Shinichi Toyooka, Takuya Fukazawa, Takafumi Kubo, Junichi Soh, Hiroaki Asano, Takayuki Muraoka, Norimitsu Tanaka, Yuho Maki, Kazuhiko Shien, Masashi Furukawa, Masakiyo Sakaguchi, Hiromasa Yamamoto, Kazunori Tsukuda, Shinichiro Miyoshi

    ACTA MEDICA OKAYAMA   68 ( 1 )   23 - 26   2014.2

     More details

    Language:English   Publisher:OKAYAMA UNIV MED SCHOOL  

    The microRNA-34s (miR-34s) have p53 response elements in their 5'-flanking regions and demonstrate tumor-suppressive functions. In malignant pleural mesothelioma (MPM), we previously reported that expression of miR-34b and miR-34c (miR-34b/c) was frequently downregulated by methylation in MPM cell lines and primary tumors. The forced overexpression of miR-34b/c showed significant antitumor effects with the induction of apoptosis in MPM cells. In this study, we examined the in vivo antitumor effects of miR-34b/c using adenovirus vector on MPM. We subcutaneously transplanted NCI-H290, a human MPM cell line, into BALB/C mice and injected adenovirus vector expressing miR-34b/c, luciferase driven by the cytomegalovirus promoter (Ad-miR-34b/c or Ad-Luc), or PBS control into tumors over 5mm in diameter. A statistically significant growth inhibition of the tumor volume was observed in the Ad-miR-34b/c group from day 6 onward compared to the Ad-Luc group. The inhibition rate of Ad-miR-34b/c, compared to the tumor volume treated with Ad-Luc, was 58.6% on day 10 and 54.7% on day13. Our results indicate that adenovirus-mediated miR-34b/c gene therapy could be useful for the clinical treatment of MPM.

    DOI: 10.18926/AMO/52140

    Web of Science

    researchmap

  • Novel Germline Mutation in the Transmembrane Domain of HER2 in Familial Lung Adenocarcinomas

    Hiromasa Yamamoto, Koichiro Higasa, Masakiyo Sakaguchi, Kazuhiko Shien, Junichi Soh, Koichi Ichimura, Masashi Furukawa, Shinsuke Hashida, Kazunori Tsukuda, Nagio Takigawa, Keitaro Matsuo, Katsuyuki Kiura, Shinichiro Miyoshi, Fumihiko Matsuda, Shinichi Toyooka

    JNCI-JOURNAL OF THE NATIONAL CANCER INSTITUTE   106 ( 1 )   djt338-djt338   2014.1

     More details

    Language:English   Publisher:OXFORD UNIV PRESS INC  

    We encountered a family of Japanese descent in which multiple members developed lung cancer. Using whole-exome sequencing, we identified a novel germline mutation in the transmembrane domain of the human epidermal growth factor receptor 2 (HER2) gene (G660D). A novel somatic mutation (V659E) was also detected in the transmembrane domain of HER2 in one of 253 sporadic lung adenocarcinomas. Because the transmembrane domain of HER2 is considered to be responsible for the dimerization and subsequent activation of the HER family and downstream signaling pathways, we performed functional analyses of these HER2 mutants. Mutant HER2 G660D and V659E proteins were more stable than wild-type protein. Both the G660D and V659E mutants activated Akt. In addition, they activated p38, which is thought to promote cell proliferation in lung adenocarcinoma. Our findings strongly suggest that mutations in the transmembrane domain of HER2 may be oncogenic, causing hereditary and sporadic lung adenocarcinomas.

    DOI: 10.1093/jnci/djt338

    Web of Science

    researchmap

  • Draft genome sequence of Clostridium botulinum type B strain Osaka05, isolated from an infant patient with botulism in Japan

    Yoshihiko Sakaguchi, Koji Hosomi, Jumpei Uchiyama, Yoshitoshi Ogura, Kaoru Umeda, Masakiyo Sakaguchi, Tomoko Kohda, Masafumi Mukamoto, Naoaki Misawa, Shigenobu Matsuzaki, Tetsuya Hayashi, Shunji Kozaki

    Genome Announcements   2 ( 1 )   e01010-e01013   2014

     More details

    © 2014 Sakaguchi et al. Clostridium botulinum strain Osaka05, which has been isolated from an infant patient with botulism in Japan, is the first strain producing botulinum neurotoxin subtype B6. Here, we report the draft genome sequence of C. botulinum Osaka05.

    DOI: 10.1128/genomeA.e01010-13

    Scopus

    researchmap

  • Inhibition of RAGE signaling through the intracellular delivery of inhibitor peptides by PEI cationization

    Endy Widya Putranto, Hitoshi Murata, Ken-Ichi Yamamoto, Ken Kataoka, Hidenori Yamada, Jun-Ichiro Futami, Masakiyo Sakaguchi, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   32 ( 4 )   938 - 944   2013.10

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    The receptor for advanced glycation end products (RAGE) is a multi-ligand cell surface receptor and a member of the immunoglobulin superfamily. RAGE is involved in a wide range of inflammatory, degenerative and hyper-proliferative disorders which span over different organs by engaging diverse ligands, including advanced glycation end products, S100 family proteins, high-mobility group protein B1 (HMGB1) and amyloid beta. We previously demonstrated that the cytoplasmic domain of RAGE is phosphorylated upon the binding of ligands, enabling the recruitment of two distinct pairs of adaptor proteins, Toll-interleukin 1 receptor domain-containing adaptor protein (TIRAP) and myeloid differentiation protein 88 (MyD88). This engagement allows the activation of downstream effector molecules, and thereby mediates a wide variety of cellular processes, such as inflammatory responses, apoptotic cell death, migration and cell growth. Therefore, inhibition of the binding of TIRAP to RAGE may abrogate intracellular signaling from ligand-activated RAGE. In the present study, we developed inhibitor peptides for RAGE signaling (RAGE-I) by mimicking the phosphorylatable cytosolic domain of RAGE. RAGE-I was efficiently delivered into the cells by polyethylenimine (PEI) cationization. We demonstrated that RAGE-I specifically bound to TIRAP and abrogated the activation of Cdc42 induced by ligand-activated RAGE. Furthermore, we were able to reduce neuronal cell death induced by an excess amount of S100B and to inhibit the migration and invasion of glioma cells in vitro. Our results indicate that RAGE-I provides a powerful tool for therapeutics to block RAGE-mediated multiple signaling.

    DOI: 10.3892/ijmm.2013.1467

    Web of Science

    researchmap

  • SARM1 and TRAF6 bind to and stabilize PINK1 on depolarized mitochondria

    Hitoshi Murata, Masakiyo Sakaguchi, Ken Kataoka, Nam-ho Huh

    MOLECULAR BIOLOGY OF THE CELL   24 ( 18 )   2772 - 2784   2013.9

     More details

    Language:English   Publisher:AMER SOC CELL BIOLOGY  

    Mutations in PTEN-induced putative kinase 1 (PINK1) or parkin cause autosomal recessive forms of Parkinson's disease. Recent work suggests that loss of mitochondrial membrane potential stabilizes PINK1 and that accumulated PINK1 recruits parkin from the cytoplasm to mitochondria for elimination of depolarized mitochondria, which is known as mitophagy. In this study, we find that PINK1 forms a complex with sterile alpha and TIR motif containing 1 (SARM1) and tumor necrosis factor receptor-associated factor 6 (TRAF6), which is important for import of PINK1 in the outer membrane and stabilization of PINK1 on depolarized mitochondria. SARM1, which is known to be an adaptor protein for Toll-like receptor, binds to PINK1 and promotes TRAF6-mediated lysine 63 chain ubiquitination of PINK1 at lysine 433. Down-regulation of SARM1 and TRAF6 abrogates accumulation of PINK1, followed by recruitment of parkin to damaged mitochondria. Some pathogenic mutations of PINK1 reduce the complex formation and ubiquitination. These results indicate that association of PINK1 with SARM1 and TRAF6 is an important step for mitophagy.

    DOI: 10.1091/mbc.E13-01-0016

    Web of Science

    researchmap

  • Tumor Necrosis Factor-alpha Down-regulates a Tumor Suppressor Gene, REIC/Dkk-3, in Normal Skin Keratinocytes

    K. Kataoka, M. Sakaguchi, H. Murata, N. Hashikawa, N. Huh

    IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY-ANIMAL   49   S43 - S43   2013.6

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:SPRINGER  

    Web of Science

    researchmap

  • DOCK7 is a critical regulator of the RAGE-Cdc42 signaling axis that induces formation of dendritic pseudopodia in human cancer cells

    Ken-Ichi Yamamoto, Hitoshi Murata, Endy Widya Putranto, Ken Kataoka, Akira Motoyama, Toshihiko Hibino, Yusuke Inoue, Masakiyo Sakaguchi, Nam-Ho Huh

    ONCOLOGY REPORTS   29 ( 3 )   1073 - 1079   2013.3

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Cellular migration is a fundamental process linked to cancer metastasis. Growing evidence indicates that the receptor for advanced glycation end products (RAGE) plays a pivotal role in this process. With regard to downstream signal transducers of RAGE, diaphanous-1 and activated small guanine nucleotide triphosphatases, Rac1 and Cdc42, have been identified. To obtain precise insight into the direct downstream signaling mechanism of RAGE, we screened for proteins interacting with the cytoplasmic domain of RAGE employing an immunoprecipitation-liquid chromatography coupled with an electrospray tandem mass spectrometry system. In the present study, we found that the cytoplasmic domain of RAGE interacted with an atypical DOCK180-related guanine nucleotide exchange factor, dedicator of cytokinesis protein 7 (DOCK7). DOCK7 bound to the RAGE cytoplasmic domain and transduced a signal to Cdc42, resulting in the formation of abundant highly branched filopodia-like protrusions, dendritic pseudopodia. Blocking of the function of DOCK7 greatly abrogated the formation of dendritic pseudopodia and suppressed cellular migration. These results indicate that DOCK7 functions as an essential and downstream regulator of RAGE-mediated cellular migration through the formation of dendritic pseudopodia.

    DOI: 10.3892/or.2012.2191

    Web of Science

    researchmap

  • S100A9 is a novel ligand of EMMPRIN that promotes melanoma metastasis

    Toshihiko Hibino, Masakiyo Sakaguchi, Shoko Miyamoto, Mami Yamamoto, Akira Motoyama, Junichi Hosoi, Tadashi Shimokata, Tomonobu Ito, Ryoji Tsuboi, Nam-Ho Huh

    Cancer Research   73 ( 1 )   172 - 183   2013.1

     More details

    Language:English  

    The calcium-binding proteins S100A8 and S100A9 can dimerize to form calprotectin, the release of which during tissue damage has been implicated in inflammation and metastasis. However, receptor(s) mediating the physiologic and pathophysiologic effects of this damage-associated "danger signal" are uncertain. In this study, searching for candidate calprotectin receptors by affinity isolation-mass spectrometry, we identified the cell surface glycoprotein EMMPRIN/BASIGIN (CD147/BSG). EMMPRIN specifically bound to S100A9 but not S100A8. Induction of cytokines and matrix metalloproteases (MMP) by S100A9 was markedly downregulated in melanoma cells by attenuation of EMMPRIN. We found that EMMPRIN signaling used the TNF receptor-associated factor TRAF2 distinct from the known S100-binding signaling pathway mediated by RAGE (AGER). S100A9 strongly promoted migration when EMMPRIN was highly expressed, independent of RAGE, whereas EMMPRIN blockade suppressed migration by S100A9. Immunohistologic analysis of melanomas revealed that EMMPRIN was expressed at both the invasive edge of lesions and the adjacent epidermis, where S100A9 was also strongly expressed. In epidermal-specific transgenic mice, tail vein-injected melanoma accumulated in skin expressing S100A9 but not S100A8. Together, our results establish EMMPRIN as a receptor for S100A9 and suggest the therapeutic use in targeting S100A9-EMMPRIN interactions. ©2012 AACR.

    DOI: 10.1158/0008-5472.CAN-11-3843

    Scopus

    PubMed

    researchmap

  • Identification of RAGE-coupled membran adaptor that modulates RAGE-triggered signaling pathway

    M. Sakaguchi, H. Murata, T. Hibino, Y. Aoyama, K. Iwatsuki, N-H. Huh

    MOLECULAR BIOLOGY OF THE CELL   24   2013

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • TRAF6 ubiquitinates and stabilizes PINK1 on the outer membrane of depolarized mitochondria through interaction with SARM1

    H. Murata, M. Sakaguchi, K. Kataoka, N-H. Huh

    MOLECULAR BIOLOGY OF THE CELL   24   2013

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Preclinical biodistribution and safety study of reduced expression in immortalized cells/Dickkopf-3-encoding adenoviral vector for prostate cancer gene therapy

    Morito Sugimoto, Masami Watanabe, Haruki Kaku, Shun-Ai Li, Hirofumi Noguchi, Hideo Ueki, Masakiyo Sakaguchi, Nam-Ho Huh, Yasutomo Nasu, Hiromi Kumon

    ONCOLOGY REPORTS   28 ( 5 )   1645 - 1652   2012.11

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    The biodistribution and safety of adenoviral vectors encoding the human REIC/Dkk-3 tumor suppressor gene (Ad-REIC) were examined in this preclinical study for in situ prostate cancer gene therapy. First, the in vitro apoptotic effects of Ad-REIC in normal and cancer cells derived from the prostate and liver were examined. Significant apoptotic effects were observed at 100 MOI (multiplicity of infection) in prostate cancer cells (LNCaP, PC3) and hepatoma cells (HEP3B and HEPG2); however, no effects were seen in normal cells. To analyze the safety of intraprostatic Ad-REIC administration, the biodistribution and histology after Ad-REIC injection were evaluated in various organs of normal male C57BL6 mice. In a supporting study, vector dissemination following intravenous injection of Ad-REIC into tail veins was determined. To evaluate whether Ad-REIC was present in the collected tissue specimens, human REIC gene detection was performed using DNA-PCR. Intraprostatic treatment administered at lower doses showed vector biodistribution into the colon, urinary bladder and prostate. At higher doses, vector dissemination was observed in tissues more distant from the prostate, including the lung, thymus, heart, liver and adrenal gland. After intravenous injection a: Ad-REIC, dissemination was observed in the liver and spleen. These results indicate that the biodistribution of Ad-REIC is determined by the dose and route of administration. Although acute inflammatory effects were observed in the prostate after intraprostatic administration at higher doses, no abnormal histological findings were noted in the other tissues, including those of intravenously treated mice. Regarding the safety of Ad-REIC administration, no deaths and no signs of toxicity or unusual behavior were observed in the mice in any treatment group. Based on these preclinical experiments, adenovirus-mediated in situ REIC/Dkk-3 gene therapy is considered to be safe for use as a treatment for human prostate cancer.

    DOI: 10.3892/or.2012.2001

    Web of Science

    researchmap

  • REIC/Dkk-3-encoding adenoviral vector as a potentially effective therapeutic agent for bladder cancer

    Takeshi Hirata, Masami Watanabe, Haruki Kaku, Yasuyuki Kobayashi, Hiroshi Yamada, Masakiyo Sakaguchi, Kohji Takei, Nam-Ho Huh, Yasutomo Nasu, Hiromi Kumon

    INTERNATIONAL JOURNAL OF ONCOLOGY   41 ( 2 )   559 - 564   2012.8

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Bladder cancer is one of the most common urogenital malignancies. The intravesical instillation of anticancer agents is an attractive strategy to treat a superficial lesion or floating/disseminated cancer cells after transurethral operation. An adenovirus carrying REIC/Dkk-3, a tumor suppressor gene (Ad-REIC), exhibits cancer-specific apoptotic effects in various types of cancer cells. The aim of the present study was to examine the potential of Ad-REIC as a therapeutic agent for bladder cancer. KK47 and RT4 human bladder cancer cells were sensitive to the Ad-REIC treatment for apoptosis induction, but some human bladder cancer cell lines (T24, J82 and TccSup) were resistant. Significant cell growth inhibition was observed when these resistant cancer cell lines were treated with Ad-REIC in a condition of floating cells, which is clinically observed after transurethral operation and becomes a cause of intravesical cancer dissemination. The therapeutic potential of Ad-REIC for the treatment of multidrug-resistant bladder cancer was investigated. The adriamycin-resistant KK47 bladder cancer cells (KK47/ADM), which also present multidrug resistance, showed induction of significant apoptosis following Ad-REIC treatment. The Ad-REIC treatment induced downregulation of P-glycoprotein in KK47/ADM cells and restored the sensitivity to doxorubicin (adriamycin). Ad-REIC suppressed P-glycoprotein expression in a c-Jun-NH2-kinase (JNK)-dependent manner. Therefore, the current study indicated two therapeutic aspects of the Ad-REIC agent against human bladder cancer cells, as an apoptosis inducer/cell growth inhibitor and as a sensitizer of chemotherapeutic agents in multidrug-resistant cancer cells. The intravesical instillation of Ad-REIC could be an attractive therapeutic method in human bladder cancer where the treatment of superficial lesions and floating/disseminated or multidrug-resistant cancer cells is necessary.

    DOI: 10.3892/ijo.2012.1503

    Web of Science

    researchmap

  • S100A8およびS100A9タンパク質の新規受容体の探索とそのがん進展における役割(Identification of novel receptors for pro-inflammatory S100A8/A9 proteins and their potential roles in tumor progression)

    阪口 政清, 日比野 利彦, 村田 等, 井上 裕介, 山本 健一, 片岡 健, 許 南浩

    日本癌学会総会記事   71回   59 - 59   2012.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Expression pattern of REIC/Dkk-3 in mouse squamous epithelia

    K. Kataoka, G. Du, N. Maehara, H. Murata, M. Sakaguchi, N. Huh

    CLINICAL AND EXPERIMENTAL DERMATOLOGY   37 ( 4 )   428 - 431   2012.6

     More details

    Language:English   Publisher:WILEY-BLACKWELL  

    The REIC/Dkk (reduced expression in immortalized cells/Dickkopf-3) gene was originally identified as a tumour-suppressor gene with reduced expression in immortalized cells, cancer-cell lines and tumour tissues. Of the four members of the Dkk family, the REIC/Dkk-3 protein is unique in terms of DNA sequence, expression profiles and biological functions. In this study, we investigated and compared the expression patterns of the REIC/Dkk-3 protein in mouse squamous epithelia. Expression of REIC/Dkk-3 in the back skin was localized to the upper layer of the interfollicular epidermis, and the inner root sheath of hair follicles. Expression of REIC/Dkk-3 was detected in the ear skin, oral mucosa, tongue, oesophagus, uterine cervix, footpad and tail skin, but not in the cornea. Interestingly, expression was localized to the upper layers of these epithelial tissues. The physiological function of REIC/Dkk-3 is still unclear, but our detailed observation highlight its unique expression pattern in squamous epithelia.

    DOI: 10.1111/j.1365-2230.2011.04301.x

    Web of Science

    researchmap

  • S100A7 promotes the migration and invasion of osteosarcoma cells via the receptor for advanced glycation end products

    Ken Kataoka, Tomoyuki Ono, Hitoshi Murata, Mika Morishita, Ken-Ichi Yamamoto, Masakiyo Sakaguchi, Nam-Ho Huh

    ONCOLOGY LETTERS   3 ( 5 )   1149 - 1153   2012.5

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Osteosarcoma is the most common malignant tumor of bone in childhood and adolescence. Despite intensive research for new therapies, the outcome in patients with metastasis remains extremely poor. S100 proteins are involved in the proliferation, cell cycle progression and metastasis of numerous malignant tumors, including osteosarcoma. In the present study, we identified S100A7 as a candidate to promote the migration of osteosarcoma cells. S100A7 promoted the migration and invasion of osteosarcoma cells as assayed in vitro. An in vitro pull-down assay revealed the binding of the recombinant S100A7 protein with its putative receptor, the receptor for advanced glycation end products (RAGE). The downregulation of RAGE by a specific siRNA markedly suppressed the migration and invasion of osteosarcoma cells. Furthermore, the matrix metalloproteinase activity of osteosarcoma cells was enhanced by S100A7 and suppressed by the downregulation of RAGE. These results indicate that S100A7 promotes the migration and invasion of osteosarcoma cells through RAGE. The S100A7-RAGE axis may thus be a new target for preventing the invasion and/or metastasis of osteosarcoma.

    DOI: 10.3892/ol.2012.612

    Web of Science

    researchmap

  • PS-109-6 REIC/Dickkopf-3 shows anti-proliferative effect for non-small cell lung cancer cells with negative phosphorylated-Akt status

    Tanaka Norimitsu, Toyooka Shinichi, Watanabe Masami, Soh Junichi, Sakaguchi Masakiyo, Tsukuda Kazunori, Asano Hiroaki, Shien Kazuhiko, Furukawa Masashi, Maki Yuho, Muraoka Takayuki, Huh Nam-ho, Nasu Yasutomo, Kumon Hiromi, Miyoshi Shinichiro

    Journal of Japan Surgical Society   113 ( 2 )   710 - 710   2012.3

     More details

    Language:English   Publisher:Japan Surgical Society (JSS)  

    CiNii Article

    CiNii Books

    researchmap

  • Partial sensitization of human bladder cancer cells to a gene-therapeutic adenovirus carrying REIC/Dkk-3 by downregulation of BRPK/PINK1

    Yu Jin, Hitoshi Murata, Masakiyo Sakaguchi, Ken Kataoka, Masami Watanabe, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    ONCOLOGY REPORTS   27 ( 3 )   695 - 699   2012.3

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    REIC/Dkk-3 is a tumor suppressor gene that was first identified as a gene downregulated in association with immortalization of normal human fibroblasts. We have demonstrated that an adenovirus carrying REIC/Dkk-3 (Ad-REIC) showed a tumor-specific killing effect on a wide range of cancers. However, some human cancers, bladder cancers in particular, are resistant to Ad-REIC. In this study, we investigated the combination effect of downregulation of BRPK/PINK1 (PINK1) and Ad-REIC on bladder cancer cells. Five bladder cancer cell lines among six cell lines examined were resistant to Ad-REIC. Among the cell lines, the resistance of two cell lines was probably due to low infection efficiency of the adenovirus. PINK1-specific siRNA remarkably downregulated Bcl-x(L) and TRAP1 proteins and upregulated BAX protein expression. Finally, downregulation of PINK1 partially sensitized the other three cell lines that were resistant to Ad-REIC. This sensitization was associated with increasing production of reactive oxygen species (ROS). These results indicate that PINK1 is one of the key molecules for the mitochondrial protection system and that PINK1 can be a new target molecule to sensitize bladder cancer cells that are resistant to Ad-REIC.

    DOI: 10.3892/or.2011.1543

    Web of Science

    researchmap

  • Preclinical Safety and Efficacy of in Situ REIC/Dkk-3 Gene Therapy for Prostate Cancer

    Keiichiro Kawauchi, Masami Watanabe, Haruki Kaku, Peng Huang, Kasumi Sasaki, Masakiyo Sakaguchi, Kazuhiko Ochiai, Nam-ho Huh, Yasutomo Nasu, Hiromi Kumon

    ACTA MEDICA OKAYAMA   66 ( 1 )   7 - 16   2012.2

     More details

    Language:English   Publisher:OKAYAMA UNIV MED SCHOOL  

    The preclinical safety and therapeutic efficacy of adenoviral vectors that express the REIC/Dkk-3 tumor suppressor gene (Ad-REIC) was examined for use in prostate cancer gene therapy. The Ad-human (h) and mouse (m) REIC were previously demonstrated to induce strong anti-cancer effects in vitro and in vivo, and we herein report the results of two in vivo studies. First, intra-tumor Ad-hREIC administration was examined for toxicity and therapeutic effects in a subcutaneous tumor model using the PC3 prostate cancer cell line. Second, intra-prostatic Ad-mREIC administration was tested for toxicity in normal mice. The whole-body and spleen weights, hematological and serum chemistry parameters, and histological evaluation of tissues from throughout the body were analyzed. Both experiments indicated that there was no significant difference in the examined parameters between the Ad-REIC-treated group and the control (PBS- or Ad-LacZ-treated) group. In the in vitro analysis using PC3 cells, a significant apoptotic effect was observed after Ad-hREIC treatment. Confirming this observation, the robust anti-tumor efficacy of Ad-hREIC was demonstrated in the in vivo subcutaneous prostate cancer model. Based on the results of these preclinical experiments, we consider the adenovirus-mediated REIC/Dkk-3 in situ gene therapy to be safe and useful for the clinical treatment of prostate cancer.

    DOI: 10.18926/AMO/48076

    Web of Science

    researchmap

  • TRAF6とSARM1によるPINK1のユビキチン化はマイトファジーの誘導に必須である

    村田等, 阪口政清, 片岡健, HUH Nam‐ho

    日本分子生物学会年会プログラム・要旨集(Web)   35th   4P-0506 (WEB ONLY)   2012

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • C型ボツリヌス毒素変換ファージのゲノム情報を基盤とした偽溶原性メカニズムの解析

    阪口義彦, OLIVA Maria A, MARTIN‐GALIANO Antonio J, ANDREU Jose M, 阪口政清, 内山淳平, 小椋義俊, 山本由弥子, 鈴木智典, 織田華絵, 津田千秋, 松崎茂展, 林哲也, 小熊惠二

    日本分子生物学会年会プログラム・要旨集(Web)   35th   WEB ONLY 3W3III-2   2012

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • 多機能受容体RAGEの下流信号伝達機構の解明(TIRAP is a critical transducer of RAGE-mediated inflammatory signaling)

    阪口 政清, 村田 等, 山本 健一, 阪口 義彦, 本山 晃, 日比野 利彦, 片岡 健, 許 南浩

    日本癌学会総会記事   70回   225 - 225   2011.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • PINK1発現抑制によるミトコンドリア機能不全はREIC/Dkk-3のアポトーシス誘導効果を増強する(Mitochondrial malfunction by PINK1 knockdown augments apoptosis induced by adenovirus carrying REIC/Dkk-3)

    村田 等, 金 玉, 阪口 政清, 片岡 健, 許 南浩

    日本癌学会総会記事   70回   51 - 51   2011.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • S100A7による骨肉腫細胞の遊走・浸潤の多機能受容体RAGEを介した制御機構(Promotion of migration and invasion of osteosarcoma cells by S100A7 through RAGE)

    片岡 健, 阪口 政清, 小野 智之, 森下 美加, 山本 健一, 村田 等, 許 南浩

    日本癌学会総会記事   70回   79 - 79   2011.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Tumor suppressor REIC/Dkk-3 interacts with the dynein light chain, Tctex-1

    Kazuhiko Ochiai, Masami Watanabe, Hideo Ueki, Peng Huang, Yasuyuki Fujii, Yasutomo Nasu, Hirofumi Noguchi, Takeshi Hirata, Masakiyo Sakaguchi, Nam-ho Huh, Yuji Kashiwakura, Haruki Kaku, Hiromi Kumon

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   412 ( 2 )   391 - 395   2011.8

     More details

    Language:English   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    REIC/Dkk-3 is a member of the Dickkopf family proteins known as Wnt-antagonists, and REIC/Dkk-3 expression is downregulated in a broad range of cancer types. REIC/Dkk-3 acts as a tumor suppressor in multiple cancer cell lines by inducing apoptosis through endoplasmic reticulum (ER) stress signaling. However, the intracellular interaction partners of REIC/Dkk-3 have not been fully elucidated. By employing yeast two-hybrid screening, we identified the human dynein light chain, Tctex-1, as a novel interaction partner of REIC/Dkk-3. We further disclosed that the interaction involves the 136-157 amino acid region of REIC/Dkk-3 by using the mammalian two-hybrid system. Interestingly, this binding region of REIC/Dkk-3 with Tctex-1 contains an amino acid sequence motif [-(E) under bar -X-(G) under bar-(R) under bar-(R) under bar -X-(H) under bar-] which was previously reported as the Tctex-1 binding domain of dynein intermediate chain (DIC). Immunocytochemistry demonstrated that both REIC/Dkk-3 and Tctex-1 were localized around the ER of human fibroblasts, and the similar distribution pattern of the proteins suggests that their interaction occurs around the ER. This is the first study showing the interaction of a Dickkopf family protein with a dynein motor complex protein. The link between REIC/Dkk-3 and Tctex-1 may be of significance for understanding the molecular functions of the proteins in ER stress signaling and intracellular dynein motor dynamics, respectively. (C) 2011 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2011.07.109

    Web of Science

    researchmap

  • Epigenetic Silencing of MicroRNA-34b/c Plays an Important Role in the Pathogenesis of Malignant Pleural Mesothelioma

    Takafumi Kubo, Shinichi Toyooka, Kazunori Tsukuda, Masakiyo Sakaguchi, Takuya Fukazawa, Junichi Soh, Hiroaki Asano, Tsuyoshi Ueno, Takayuki Muraoka, Hiromasa Yamamoto, Yasutomo Nasu, Takumi Kishimoto, Harvey I. Pass, Hideki Matsui, Nam-ho Huh, Shinichiro Miyoshi

    CLINICAL CANCER RESEARCH   17 ( 15 )   4965 - 4974   2011.8

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    Purpose: Malignant pleural mesothelioma (MPM) is an aggressive tumor with a dismal prognosis. Unlike other malignancies, TP53 mutations are rare in MPM. Recent studies have showed that altered expression of microRNA (miRNA) is observed in human malignant tumors. In this study, we investigated the alterations of miR-34s, a direct transcriptional target of TP53, and the role of miR-34s on the pathogenesis of MPM.
    Experimental Design: Aberrant methylation and expression of miR-34s were examined in MPM cell lines and tumors. miR-34b/c was transfected to MPM cells to estimate the protein expression, cell proliferation, invasion, and cell cycle.
    Results: Aberrant methylation was present in 2 (33.3%) of 6 MPM cell lines and 13 (27.7%) of 47 tumors in miR-34a and in all 6 MPM cell lines (100%) and 40 (85.1%) of 47 tumors in miR-34b/c. Expression of miR-34a and 34b/c in all methylated cell lines was reduced and restored with 5-aza-2&apos;-deoxycytidine treatment. Because epigenetic silencing was the major event in miR-34b/c, we investigated the functional role of miR-34b/c in MPM. miR-34b/c-transfected MPM cells with physiologic miR-34b/c expression exhibited antiproliferation with G(1) cell cycle arrest and suppression of migration, invasion, and motility. The forced overexpression of miR-34b/c, but not p53, showed a significant antitumor effect with the induction of apoptosis in MPM cells.
    Conclusions: We show that the epigenetic silencing of miR-34b/c by methylation is a crucial alteration and plays an important role in the tumorigenesis of MPM, suggesting potential therapeutic options for MPM. Clin Cancer Res; 17(15); 4965-74. (C)2011 AACR.

    DOI: 10.1158/1078-0432.CCR-10-3040

    Web of Science

    researchmap

  • TIRAP, an Adaptor Protein for TLR2/4, Transduces a Signal from RAGE Phosphorylated upon Ligand Binding

    Masakiyo Sakaguchi, Hitoshi Murata, Ken-ichi Yamamoto, Tomoyuki Ono, Yoshihiko Sakaguchi, Akira Motoyama, Toshihiko Hibino, Ken Kataoka, Nam-ho Huh

    PLOS ONE   6 ( 8 )   23132 - 23132   2011.8

     More details

    Language:English   Publisher:PUBLIC LIBRARY SCIENCE  

    The receptor for advanced glycation end products (RAGE) is thought to be involved in the pathogenesis of a broad range of inflammatory, degenerative and hyperproliferative diseases. It binds to diverse ligands and activates multiple intracellular signaling pathways. Despite these pivotal functions, molecular events just downstream of ligand-activated RAGE have been surprisingly unknown. Here we show that the cytoplasmic domain of RAGE is phosphorylated at Ser391 by PKCf upon binding of ligands. TIRAP and MyD88, which are known to be adaptor proteins for Toll-like receptor-2 and -4 (TLR2/4), bound to the phosphorylated RAGE and transduced a signal to downstream molecules. Blocking of the function of TIRAP and MyD88 largely abrogated intracellular signaling from ligand-activated RAGE. Our findings indicate that functional interaction between RAGE and TLRs coordinately regulates inflammation, immune response and other cellular functions.

    DOI: 10.1371/journal.pone.0023132

    Web of Science

    researchmap

  • Markedly elevated serum levels of calcium-binding S100A8/A9 proteins in psoriatic arthritis are due to activated. monocytes/macrophages

    Seiko Aochi, Kazuhide Tsuji, Masakiyo Sakaguchi, Namho Huh, Tatsuya Tsuda, Kiyofumi Yamanishi, Mayumi Komine, Keiji Iwatsuki

    JOURNAL OF THE AMERICAN ACADEMY OF DERMATOLOGY   64 ( 5 )   879 - 887   2011.5

     More details

    Language:English   Publisher:MOSBY-ELSEVIER  

    Background: Serum levels of S100A8/A9 may correlate with disease activity in psoriasis.
    Objective: We sought to elucidate the association of serum levels of S100A8/A9 heterodimers with the clinical subtypes of psoriasis and the major cell source.
    Methods: Serum samples were collected from patients with psoriasis vulgaris (n = 30), psoriatic arthritis (PA) (n = 16),. pustular psoriasis (n = 24), and atopic dermatitis (n = 14) and from healthy control subjects (n = 21). Serum concentrations of S100A8/A9 were measured, and the expression levels were examined in psoriatic lesions. The messenger RNA levels were quantified in circulating monocytes and neutrophils.
    Results: Serum levels of S100A8/A9 were significantly increased in all subtypes of psoriasis as compared with healthy controls and atopic dermatitis. Among the psoriatic subtypes, PA and pustular psoriasis showed remarkably high concentrations of S100A8/A9 heterodimers. The higher serum levels were associated with the presence of articular symptoms, but not significantly correlated with body surface areas of psoriatic lesions. S100A8 was expressed by both keratinocytes and infiltrating mononuclear cells, whereas S100A9 was predominantly expressed by neutrophils. The expression levels of S100A8 and S100A9 messenger RNA in monocytes were increased by approximately 2.25- and 1.91-fold in PA, respectively, whereas no significant increase was observed in psoriasis vulgaris and pustular psoriasis.
    Limitations: Difficulty in acquisition of clinical and laboratory samples in untreated patients, and of a sufficient number of subjects, were limitations.
    Conclusions: Although serum levels of S100A8/A9 were increased in all types of psoriasis examined, patients with PA had higher levels of S100A8/A9, probably because of an activated monocyte/macrophage system. (J Am Acad Dermatol 2011;64:879-87.)

    DOI: 10.1016/j.jaad.2010.02.049

    Web of Science

    researchmap

  • Multiple functions of PINK1 at different intracellular locations Beyond neurodegenerative diseases

    Hitoshi Murata, Masakiyo Sakaguchi, Ken Kataoka, Nam-ho Huh

    CELL CYCLE   10 ( 10 )   1518 - 1519   2011.5

     More details

    Language:English   Publisher:LANDES BIOSCIENCE  

    DOI: 10.4161/cc.10.10.15445

    Web of Science

    researchmap

  • Intraperitoneal administration of an adenovirus vector carrying REIC/Dkk-3 suppresses peritoneal dissemination of scirrhous gastric carcinoma

    Swe Swe Than, Ken Kataoka, Masakiyo Sakaguchi, Hitoshi Murata, Fernando Abarzua, Chika Taketa, Gang Du, Masakazu Yashiro, Kazuyoshi Yanagihara, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    ONCOLOGY REPORTS   25 ( 4 )   989 - 995   2011.4

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    Expression levels of the novel tumor suppressor gene REIC/Dkk-3 are reduced in many human cancers. We have previously showed that an adenovirus vector carrying REIC/Dkk-3 (Ad-REIC) induced apoptosis of cancer cells selectively and exerted bystander antitumor effects via ER stress. We examined possible effects of Ad-REIC in a peritoneal dissemination model of scirrhous gastric carcinoma (SGC). Among various types of gastric cancer, SGC continues to be associated with the worst prognosis due to a high incidence of metastases in the peritoneal cavity. We found that a single intraperitoneal injection of Ad-REIC suppressed tumor dissemination and disease progression. Immunomodulation by Ad-REIC led to recruitment of natural killer cells inside tumor nodules. We conclude that Ad-REIC gene therapy may be a potential tool in combinatorial approaches to achieve curative effects in SGC.

    DOI: 10.3892/or.2011.1149

    Web of Science

    researchmap

  • Identification of EMMPRIN as a novel receptor for S100A9: Its involvement in melanoma metastasis

    T. Hibino, S. Miyamoto, M. Sakaguchi, A. Motoyama, M. Yamamoto, R. Tsuboi, N. Huh

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   131   S126 - S126   2011.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:NATURE PUBLISHING GROUP  

    Web of Science

    researchmap

  • Expression of REIC/Dkk-3 in normal and hyperproliferative epidermis

    Gang Du, Ken Kataoka, Masakiyo Sakaguchi, Fernando Abarzua, Swe Swe Than, Hiroyuki Sonegawa, Teruhiko Makino, Tadamichi Shimizu, Nam-Ho Huh

    EXPERIMENTAL DERMATOLOGY   20 ( 3 )   273 - 277   2011.3

     More details

    Language:English   Publisher:WILEY-BLACKWELL  

    Dickkopf (Dkk) family members are known as Wnt modulators involved in the development, cell growth/differentiation and cancer. REIC/Dkk-3, which does not interfere with Wnt signalling, has been proposed to be a tumor suppressor gene, but its physiological function has remained unclear. In this study, we analysed the expression of REIC/Dkk-3 in normal interfollicular epidermis (IFE) and hyperproliferative epidermis. REIC/Dkk-3 was expressed in human and mouse IFE, being localized at the interface of upper spinous layer and granular layer. Skin cancer cell lines lost REIC/Dkk-3 expression as reported previously. When we analysed patient samples, REIC/Dkk-3 expression was down-regulated in the hyperproliferative epidermis including skin cancers and non-cancerous proliferative diseases. REIC/Dkk-3 expression was also suppressed in the regenerative and inflammative epidermis of model mice. These findings will certainly contribute to the extension of studies on REIC/Dkk-3.

    DOI: 10.1111/j.1600-0625.2010.01244.x

    Web of Science

    researchmap

  • REIC/Dkk-3の扁平上皮における発現とその制御因子の探索

    片岡 健, 阪口 政清, 杜 剛, 前原 奈都美, 村田 等, 許 南浩

    組織培養研究   30 ( 1 )   99 - 99   2011.3

     More details

    Language:Japanese   Publisher:日本組織培養学会  

    researchmap

  • A New Cytosolic Pathway from a Parkinson Disease-associated Kinase, BRPK/PINK1 ACTIVATION OF AKT VIA MTORC2

    Hitoshi Murata, Masakiyo Sakaguchi, Yu Jin, Yoshihiko Sakaguchi, Jun-ichiro Futami, Hidenori Yamada, Ken Kataoka, Nam-ho Huh

    JOURNAL OF BIOLOGICAL CHEMISTRY   286 ( 9 )   7182 - 7189   2011.3

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Accumulating evidence indicates that dysfunction of mitochondria is a common feature of Parkinson disease. Functional loss of a familial Parkinson disease-linked gene, BRPK/PINK1 (PINK1), results in deterioration of mitochondrial functions and eventual neuronal cell death. A mitochondrial chaperone protein has been shown to be a substrate of PINK1 kinase activity. In this study, we demonstrated that PINK1 has another action point in the cytoplasm. Phosphorylation of Akt at Ser-473 was enhanced by overexpression of PINK1, and the Akt activation was crucial for protection of SH-SY5Y cells from various cytotoxic agents, including oxidative stress. Enhanced Akt phosphorylation was not due to activation of phosphatidylinositol 3-kinase but due to activation of mammalian target of rapamycin complex 2 (mTORC2) by PINK1. Rictor, a specific component of mTORC2, was phosphorylated by overexpression of PINK1. Furthermore, overexpression of PINK1 enhanced cell motility. These results indicate that PINK1 exerts its cytoprotective function not only in mitochondria but also in the cytoplasm through activation of mTORC2.

    DOI: 10.1074/jbc.M110.179390

    Web of Science

    researchmap

  • A novel tumor suppressor, REIC/Dkk-3 gene identified by our in vitro transformation model of normal human fibroblasts works as a potent therapeutic anti-tumor agent

    Masakiyo Sakaguchi, Nam-Ho Huh, Masayoshi Namba

    Advances in Experimental Medicine and Biology   720   209 - 215   2011

     More details

    Language:English  

    Reduced Expression in Immortalized Cell (REIC) was cloned by subtractive hybridization method as a gene whose expression is reduced in many human immortalized and neoplastic tumor cells. The REIC, when over-expressed by an adenovirus (Ad-REIC), exhibited a dramatic therapeutic effect on a wide variety of human cancers through a mechanism triggered by ER-stress-mediated JNK activation. In addition to this direct effect on cancer cells, Ad-REIC exerted another cytotoxicity on human cancers, an indirect host-mediated effect due to overproduction of IL-7 by mis-targeted normal cells. This "one-bullet two-arms" finding may lead to a powerful new therapeutic approach to the treatment of human cancers. © 2011 Springer Science+Business Media, LLC.

    DOI: 10.1007/978-1-4614-0254-1_17

    Scopus

    PubMed

    researchmap

  • 正常皮膚におけるREIC/Dkk‐3の発現制御因子の検索

    前原奈都美, 片岡健, DU Gang, 村田等, 山本健一, 阪口政清, HUH Nam‐Ho

    日本分子生物学会年会プログラム・要旨集(Web)   34th   1P-0254 (WEB ONLY)   2011

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • S100タンパク質受容体の結合解析

    森下美加, 村田等, 山本健一, 片岡健, 阪口政清, HUH Nam‐Ho

    日本分子生物学会年会プログラム・要旨集(Web)   34th   4P-0293 (WEB ONLY)   2011

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • Expression pattern of REIC/Dkk-3 in various cell types and the implications of the soluble form in prostatic acinar development

    Kai Zhang, Masami Watanabe, Yuji Kashiwakura, Shun-Al Li, Kohei Edamura, Peng Huang, Ken Yamaguchi, Yasutomo Nasu, Yasuyuki Kobayashi, Masakiyo Sakaguchi, Kazuhiko Ochiai, Hiroshi Yamada, Kohji Takei, Hideo Ueki, Nam-Ho Huh, Ming Li, Haruki Kaku, Yanqun Na, Hiromi Kumon

    INTERNATIONAL JOURNAL OF ONCOLOGY   37 ( 6 )   1495 - 1501   2010.12

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    The tumor suppressor REIC/Dkk-3 is a secretory protein which was originally identified to be downregulated in human immortalized cells In the present study, we investigated the expression pattern of REIC/Dkk-3 in various cell types to characterize its physiological functions We first examined the expression level of REIC/Dkk-3 in a broad range of cancer cell types and confirmed that it was significantly downregulated in all of the cell types We also examined the tissue distribution pattern in a variety of normal mouse organs Ubiquitous REIC/Dkk-3 protein expression was observed in the organs The expression was abundant in the liver, heart and brain tissue, but was absent in the spleen and peripheral blood mononuclear cells The immunohistochemical analyses revealed that the subcellular localization of REIC/Dkk-3 had a punctate pattern around the nucleus, indicating its association with secretory vesicles In cancer cells stably transfected with REIC/Dkk-3 the protein was predominantly localized to the endoplasmic reticulum (ER) under observation with confocal microscopy Because REIC/ Dkk-3 was found to be abundantly expressed in the acinar epithelial cells of the mouse prostate, we analyzed the effects of recombinant REIC/Dkk-3 protein on the acinar morphogenesis of RWPE-1 cells, which are derived from human normal prostate epithelium Statistically significant acinar growth was observed in the culture condition with 10 mu g/m1 REIC/Dkk-3 protein, implicating the soluble form m prostatic acinar development Current results suggest that REIC/Dkk-3 may play a role in regulating the morphological process of normal tissue architecture through an autocrine and/or paracrine manner

    DOI: 10.3892/ijo_00000802

    Web of Science

    researchmap

  • KLF11とKLF15によるUCP1の発現制御

    山本 健一, 阪口 政清, 片岡 健, 許 南浩

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集   83回・33回   2P - 0822   2010.12

     More details

    Language:Japanese   Publisher:(公社)日本生化学会  

    researchmap

  • BRPK/PINK1のmTORC2経路活性化を介したがん進展への寄与(BRPK/PINK1 promotes tumor progression through activation of mTORC2 pathway)

    村田 等, 阪口 政清, 金 玉, 片岡 健, 許 南浩

    日本生化学会大会・日本分子生物学会年会合同大会講演要旨集   83回・33回   4P - 0934   2010.12

     More details

    Language:English   Publisher:(公社)日本生化学会  

    researchmap

  • Internalization of REIC/Dkk-3 protein by induced pluripotent stem cell-derived embryoid bodies and extra-embryonic tissues

    Ken Kataoka, Masakiyo Sakaguchi, Kun Peng Li, Chika Taketa, Ken-Ichi Yamamoto, Gang Du, Hiroaki Funahashi, Hitoshi Murata, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   26 ( 6 )   853 - 859   2010.12

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    REIC/Dkk-3 was first identified as a down-regulated gene in a number of human immortalized cells and human tumor-derived cell lines. Overexpression of the REIC/Dkk-3 gene using an adenovirus vector (Ad-REIC) has showed a potent selective therapeutic effect on various human cancers through induction of ER stress. Furthermore, we recently showed that Ad-REIC has an indirect host-mediated anti-tumor activity by induction of IL-7. However, the physiological function of REIC/Dkk-3 is still unclear. As a first step to study the possible receptor(s) for secreted REIC/Dkk-3, we analyzed the internalization of Cy3-labeled recombinant REIC/Dkk-3 protein. Among the cell lines screened, mouse induced pluripotent stem (iPS) cells showed a unique pattern of internalization. The internalization was observed in peripheral cells of spherical colonies formed spontaneously, but not in undifferentiated iPS cells. When we analyzed embryoid bodies (EBs) derived from iPS cells, REIC/Dkk-3 protein was internalized specifically by differentiated cells located at the periphery of EBs. Interestingly, Dkk-1 was internalized by undifferentiated cells at the center of the EBs. When developmental tissue was analyzed, internalization of REIC/Dkk-3 protein was strictly limited to extra-embryonic tissue, such as the trophectoderm layer of 4.5 days post-coitus (dpc) blastocysts and the chorionic membrane at 16.5 dpc. The mechanism of the internalization was confirmed to be endocytosis. These findings will contribute to knowledge on the interaction of REIC/Dkk-3 with a possible receptor(s).

    DOI: 10.3892/ijmm_00000534

    Web of Science

    researchmap

  • Transcriptional regulation of a brown adipocyte-specific gene, UCP1, by KLF11 and KLF15

    Ken-ichi Yamamoto, Masakiyo Sakaguchi, Reinhold J. Medina, Aya Niida, Yoshihiko Sakaguchi, Masahiro Miyazaki, Ken Kataoka, Nam-ho Huh

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   400 ( 1 )   175 - 180   2010.9

     More details

    Language:English   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Several growth factors and transcription factors have been reported to play important roles in brown adipocyte differentiation and modulation of thermogenic gene expression, especially the expression of UCP1. In this study, we focused on KLF11 and KLF15, which were expressed highly in brown adipose tissue. Our data demonstrated that KLF11 and KLF15 interacted directly with the UCP1 promoter using GC-box and GT-boxes, respectively. Co-transfection of KLF11 and KLF15 in the mesenchymal stem cell line muBM3.1 during brown adipocyte differentiation enhanced the expression level of UCP1. KLF11, but not KLF15, was essential for UCP1 expression during brown adipocyte differentiation of muBM3.1. (C) 2010 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2010.08.039

    Web of Science

    researchmap

  • 新規がん抑制遺伝子REIC/Dkk-3の増殖性皮膚疾患での発現減少(Reduced expression of REIC/Dkk-3, a putative tumor suppressor gene, in hyperproliferative epidermis)

    杜 剛, 片岡 健, 阪口 政清, 牧野 輝彦, 清水 忠道, 許 南浩

    日本癌学会総会記事   69回   316 - 316   2010.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • アデノウイルス受容体(CAR;CXADR)の固形癌細胞の増殖制御における役割の解析(Biological role of Coxsackievirus and Adenovirus Receptor (CXADR) in cancer cell proliferation)

    小屋 恵理子, 阪口 政清, 松井 誠, 冨田 勇樹, 許 南浩, 近藤 英作

    日本癌学会総会記事   69回   268 - 268   2010.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • スキルス胃がん腹膜播種へのREIC/Dkk-3遺伝子治療の効果(REIC/Dkk-3 gene therapy suppresses peritoneal dissemination of scirrhous gastric carcinoma)

    片岡 健, タン・スエスエ, 阪口 政清, アバルスァ・フェルナンド, 村田 等, 八代 正和, 那須 保友, 公文 裕巳, 許 南浩

    日本癌学会総会記事   69回   252 - 253   2010.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • マウス骨髄間葉系幹細胞の褐色脂肪分化をKLF11およびKLF15は促進する

    片岡健, 山本健一, 阪口政清, HUH Nam‐ho

    硬組織再生生物学会学術大会・総会プログラム・抄録集   19th   40   2010.7

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • Down-regulation of BiP/GRP78 sensitizes resistant prostate cancer cells to gene-therapeutic overexpression of REIC/Dkk-3

    Ryuta Tanimoto, Masakiyo Sakaguchi, Fernando Abarzua, Ken Kataoka, Kaoru Kurose, Hitoshi Murata, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF CANCER   126 ( 7 )   1562 - 1569   2010.4

     More details

    Language:English   Publisher:JOHN WILEY & SONS INC  

    We have recently shown that an adenovirus carrying REIC/Dkk-3 (Ad-REIC) exhibits a potent tumor-specific cell-killing function for various human cancers. It has also become evident that some human cancers are resistant to Ad-REIC-induced apoptosis. The aim of the present study was to determine the molecular mechanisms of resistance to Ad-REIC. First, we isolated resistant clones from a human prostate cancer cell line, PC3, after repeated exposure to Ad-REIC. Infection efficiency of the adenovirus vector and expression level of REIC/Dkk-3 in the resistant clones were similar to those in the parental PC3 cells. By screening for alteration in levels and functional status of proteins involved in Ad-REIC-induced apoptosis, we found that BiP/GRP78, an ER-residing chaperone protein, was expressed at higher levels consistently among resistant cells. Expression levels of BiP and rates of apoptosis induced by Ad-REIC were inversely correlated. Down-regulation of BiP with siRNA sensitized the resistant cells to Ad-REIC in vivo as well as in culture. These results indicate that BiP is a major determinant of resistance to Ad-REIC-induced apoptosis. Thus BiP is useful for diagnosis of inherent and acquired resistance of cancers and also as a target molecule to overcome resistance to the gene therapeutic Ad-REIC.

    DOI: 10.1002/ijc.24764

    Web of Science

    researchmap

  • 分化誘導したマウスES細胞及びiPS細胞におけるCy3標識REIC/Dkk‐3タンパク質のエンドサートーシスによる取り込み

    片岡健, 阪口政清, LI Kun Peng, DU Gang, 武田千佳, 舟橋弘晃, 村田等, HUH Nam‐Ho

    組織培養研究   29 ( 1 )   74   2010.3

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • 褐色脂肪細胞分化におけるKLF11とKLF15の役割

    山本健一, 阪口政清, MEDINA Reinhold, 新居田彩, 宮崎正博, 片岡健, HUH Nam‐Ho

    組織培養研究   29 ( 1 )   82   2010.3

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • 関節症性乾癬におけるバイオマーカーとしての血清S100A8/A9蛋白

    青地聖子, 辻和英, 阪口政清, 許南浩, 津田達也, 山西清文, 小宮根真弓, 岩月啓氏

    日本皮膚科学会雑誌   120 ( 3 )   691   2010.3

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • S100A11, a dual growth regulator of epidermal keratinocytes.

    Sakaguchi M, Huh NH

    Amino Acids   1111 - 1111   2010

     More details

  • BRPK/PINK1のmTORC2経路活性化を介したがん浸潤への寄与(BRPK/PINK1 enhances invasiveness of cancer cells trough activation of mTORC2 pathway)

    村田 等, 阪口 政清, 片岡 健, 許 南浩

    日本癌学会総会記事   68回   205 - 205   2009.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Overexpression of REIC/Dkk-3 in Normal Fibroblasts Suppresses Tumor Growth via Induction of Interleukin-7

    Masakiyo Sakaguchi, Ken Kataoka, Fernando Abarzua, Ryuta Tanimoto, Masami Watanabe, Hitoshi Murata, Swe Swe Than, Kaoru Kurose, Yuji Kashiwakura, Kazuhiko Ochiai, Yasutomo Nasu, Hiromi Kumon, Nam-ho Huh

    JOURNAL OF BIOLOGICAL CHEMISTRY   284 ( 21 )   14236 - 14244   2009.5

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    We previously showed that the tumor suppressor gene REIC/Dkk-3, when overexpressed by an adenovirus (Ad-REIC), exhibited a dramatic therapeutic effect on human cancers through a mechanism triggered by endoplasmic reticulum stress. Adenovirus vectors show no target cell specificity and thus may elicit unfavorable side effects through infection of normal cells even upon intra-tumoral injection. In this study, we examined possible effects of Ad-REIC on normal cells. We found that infection of normal human fibroblasts (NHF) did not cause apoptosis but induced production of interleukin (IL)-7. The induction was triggered by endoplasmic reticulum stress and mediated through IRE1 alpha, ASK1, p38, and IRF-1. When Ad-REIC-infected NHF were transplanted in a mixture with untreated human prostate cancer cells, the growth of the cancer cells was significantly suppressed. Injection of an IL-7 antibody partially abrogated the suppressive effect of Ad-REIC-infected NHF. These results indicate that Ad-REIC has another arm against human cancer, an indirect host-mediated effect because of overproduction of IL-7 by mis-targeted NHF, in addition to its direct effect on cancer cells.

    DOI: 10.1074/jbc.M808002200

    Web of Science

    researchmap

  • BRPK/PINK1のmTORC2活性化を介したがん進展への寄与

    村田 等, 阪口 政清, 片岡 健, 許 南浩

    組織培養研究   28 ( 1 )   66 - 66   2009.3

     More details

    Language:Japanese   Publisher:日本組織培養学会  

    researchmap

  • Immunological aspects of REIC/Dkk-3 in monocyte differentiation and tumor regression

    Masami Watanabe, Yuji Kashiwakura, Peng Huang, Kazuhiko Ochiai, Junichiro Futami, Shun-Ai Li, Munenori Takaoka, Yasutomo Nasu, Masakiyo Sakaguchi, Nam-Ho Huh, Hiromi Kumon

    INTERNATIONAL JOURNAL OF ONCOLOGY   34 ( 3 )   657 - 663   2009.3

     More details

    Language:English   Publisher:SPANDIDOS PUBL LTD  

    The REIC/Dkk-3 gene has been reported to be a tumor suppressor and the expression is significantly down-regulated in a broad range of cancer cell types. The protein is secretory, but the physiological function remains unclear. This study demonstrated that recombinant REIC/Dkk-3 protein induced the differentiation of human CD14(+) monocytes into a novel cell type ((REIC/Dkx-3)Mo). (REIC/Dkk-3)Mo resembles immature dendritic cells generated with IL-4 and GM-CSF. Both these cell populations exhibit similar proportions of CD11c(+), CD40(+), CD86(+) and HLA-DR(+) cells and endocytic capacity, but (REIC/Dkk-3)Mo is negative for CD1a antigen. An analysis of the signal transducers and activators of transcription (STAT) pathways revealed that REIC/Dkk-3 induces phosphorylation of STAT 1 and STAT 3. Furthermore, intratumoral administration of REIC/Dkk-3 protein significantly suppressed tumor growth with CD11c(+) and CD8(+) (dendritic and killer T cell marker, respectively) cell accumulation and enhanced anticancer cytolytic activity of splenocytes. These data indicated a cytokine-like role of REIC/Dkk-3 protein in monocyte differentiation that might be exploited therapeutically.

    DOI: 10.3892/ijo_00000191

    Web of Science

    researchmap

  • 分化したマウスES細胞およびiPS細胞のCy3標識REIC/Dkk‐3タンパク質の取り込み

    片岡健, 阪口政清, 李坤鵬, 武田千佳, 山本健一, 許南浩

    再生医療   8   223   2009.2

     More details

    Language:Japanese  

    J-GLOBAL

    researchmap

  • REIC/Dkk-3 overexpression downregulates P-glycoprotein in multidrug-resistant MCF7/ADR cells and induces apoptosis in breast cancer

    K. Kawasaki, M. Watanabe, M. Sakaguchi, Y. Ogasawara, K. Ochiai, Y. Nasu, H. Doihara, Y. Kashiwakura, N-h Huh, H. Kumon, H. Date

    CANCER GENE THERAPY   16 ( 1 )   65 - 72   2009.1

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    The overexpression of reduced expression in immortalized cells (REIC)/Dickkopf-3 (Dkk-3), a tumor suppressor gene, induced apoptosis in human prostatic and testicular cancer cells. The aim of this study is to examine the potential of REIC/Dkk-3 as a therapeutic target against breast cancer. First, the in vitro apoptotic effect of Ad-REIC treatment was investigated in breast cancer cell lines and the adenovirus-mediated overexpression of REIC/Dkk-3 was thus found to lead to apoptotic cell death in a c-Jun-NH(2)-kinase (JNK) phosphorylaion-dependent manner. Moreover, an in vivo apoptotic effect and MCF/Wt tumor growth inhibition were observed in the mouse model after intratumoral Ad-REIC injection. As multidrug resistance (MDR) is a major problem in the chemotherapy of progressive breast cancer, the in vitro effects of Ad-REIC treatment were investigated in terms of the sensitivity of multidrug-resistant MCF7/ADR cells to doxorubicin and of the P-glycoprotein expression. Ad-REIC treatment in MCF7/ADR cells also downregulated P-glycoprotein expresssion through JNK activation, and sensitized its drug resistance against doxorubicin. Therefore, not only apoptosis induction but also the reversal of anticancer drug resistance was achieved using Ad-REIC. We suggest that REIC/Dkk-3 is a novel target for breast cancer treatment and that Ad-REIC might be an attractive agent against drug-resistant cancer in combination with conventional antineoplastic agents.

    DOI: 10.1038/cgt.2008.58

    Web of Science

    researchmap

  • REIC/Dkk-3 in normal fibroblasts suppresses tumor growth via induction of interleukin-7.

    Chen J, Watanabe M, Huang P, Sakaguchi M, Ochiai K, Nasu Y, Ouchida M, Huh NH, Shimizu K, Kashiwakura Y, Kaku H, Kumon H

    Int J Mol Med   24 ( 6 )   789 - 794   2009

     More details

  • Suppression of Carbon Tetrachloride-Induced Liver Fibrosis by Transplantation of a Clonal Mesenchymal Stem Cell Line Derived From Rat Bone Marrow

    Marhaen Hardjo, Masahiro Miyazaki, Masakiyo Sakaguchi, Takuro Masaka, Sukaeni Ibrahim, Ken Kataoka, Narn-ho Huh

    CELL TRANSPLANTATION   18 ( 1 )   89 - 99   2009

     More details

    Language:English   Publisher:COGNIZANT COMMUNICATION CORP  

    Transplantation of hepatocytes or bone marrow-derived cells has been shown to ameliorate liver fibrosis in animal models, but no direct comparison of relative efficiency has been made. The aim of this study was to compare the efficiency of a bone marrow-derived clonal mesenchymal stem cell line established by LIS (rBM25/S3) with that of its adipogenic or hepatogenic differentiation derivative for Suppression of rat liver fibrosis. After induction of differentiation of rBM25/S3 cells into adipogenic or hepatogenic cells in Culture, we intrasplenically transplanted the three types of cells into rats (3 x 10(7) cells/rat) before and 4 weeks after initiation of carbon tetrachloride treatment (1 ml/kg body weight twice a week for 8 weeks) to induce liver fibrosis. Undifferentiated rBM25/S3 cells were the most effective for suppression of liver fibrosis, followed by the adipogenic cells and hepatogenic cells. Expression levels of MMP-2 and MMP-9 were also highest in undifferentiated rBM25/S3 cells. These results indicate that bone marrow-derived clonal mesenchymal stem cell lines are useful for further mechanistic studies on cell-mediated suppression of liver fibrosis and that such cell lines will provide information on an appropriate cell source for transplantation therapy for cirrhosis.

    DOI: 10.3727/096368909788237140

    Web of Science

    researchmap

  • Mechanistic Analysis of Resistance to REIC/Dkk-3-induced Apoptosis in Human Bladder Cancer Cells

    Tomoko Kobayashi, Masakiyo Sakaguchi, Ryuta Tanimoto, Fernando Abarzua, Mikiro Takaishi, Haruki Kaku, Ken Kataoka, Takashi Saika, Yasutomo Nasu, Masahiro Miyazaki, Hiromi Kumon, Nam-ho Huh

    ACTA MEDICA OKAYAMA   62 ( 6 )   393 - 401   2008.12

     More details

    Language:English   Publisher:OKAYAMA UNIV MED SCHOOL  

    We have recently shown that a new therapeutic modality using the REIC/Dkk-3 gene (Ad-REIC) is effective against various human cancers, including those of prostate, testis and breast origins. The aim of the present study was to examine the sensitivity of bladder cancers to Ad-REIC and to clarify the molecular mechanisms that determine sensitivity/resistance. We found that 2 human bladder cancer cell lines, T24 and J82, are resistant to Ad-REIC. In T24 and J82 cells, the ER stress response and activation of JNK were observed in a manner similar to that in the sensitive PC3 cells. Translocation of Bax to mitochondria occurred in PC3 cells but not in T24 and J82 cells. Bcl-2 was remarkably overexpressed in T24 and J82 compared with the expression levels in sensitive cell lines. Treatment of T24 and J82 cells with a Bcl-2 inhibitor sensitized the cells to Ad-REIC-induced apoptosis. The results indicate that some human bladder cancers are resistant to apoptosis induced by overexpression of REIC/Dkk-3, which is at least in part due to up-regrulation of Bcl-2. These results provide a basis for possible use of Bcl-2 as a marker of sensitive cancers and to try to sensitize resistant cancers to Ad-REIC by down-regrulation of Bcl-2.

    DOI: 10.18926/AMO/30945

    Web of Science

    researchmap

  • Down-regulation of Inhibition of Differentiation-1 via Activation of Activating Transcription Factor 3 and Smad Regulates REIC/Dickkopf-3-Induced Apoptosis

    Yuji Kashiwakura, Kazuhiko Ochiai, Masami Watanabe, Fernando Abarzua, Masakiyo Sakaguchi, Munenori Takaoka, Ryuta Tanimoto, Yasutomo Nasu, Nam-ho Hub, Hiromi Kumon

    CANCER RESEARCH   68 ( 20 )   8333 - 8341   2008.10

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    REIC/Dickkopf-3 (Dkk-3), a tumor suppressor gene, has been investigated in gene therapy studies. Our previous study suggested that REIC/Dkk-3-induced apoptosis mainly resulted from phosphorylation of c-Jun-NH2 kinase (JNK) in prostate cancer cells. However, the precise mechanisms, especially the molecular mechanisms regulating JNK phosphorylation, remain unclear. In this study, we investigated the mechanisms participating in JNK phosphorylation in the context of a refractory cancer disease, malignant mesothelioma (MM). Adenovirus-mediated overexpression of REIC/Dkk-3 induced apoptosis mainly through JNK activation in immortalized MM cells (211H cells). Interestingly, transcriptional down-regulation of inhibition of differentiation-1 (Id-1) was detected in REIC/Dkk-3-overexpressed 211H cells. Moreover, restoration of Id-1 expression antagonized REIC/Dkk-3-induced JNK phosphorylation and apoptosis. Mutagenesis experiments with the 2.1-kb human Id-1 promoter revealed that activating transcription factor 3 (ATF3) and Smad interaction, with their respective binding motifs, was essential for REIC/Dkk-3-mediated suppression of Id-1 promoter activity. ATF3 activation was probably induced by endoplasmic reticulum stress. Finally, we showed strong antitumor effects from REIC/Dkk-3 gene transfer into the pleural cavity in an orthotopic MM mouse model. Relative to control tumor tissue, REIC/Dkk-3-treated tumor tissue showed down-regulated expression of Id-1 mRNA, enhanced expression of phosphorylated JNK, and an increased number of apoptotic cells. In summary, we first showed that both ATF3 and Smad were crucially and synergistically involved in down-regulation of Id-1, which regulated JNK phosphorylation in REIC/Dkk-3-induced apoptosis. Thus, gene therapy with REIC/Dkk-3 may be a promising therapeutic tool for MM. [Cancer Res 2008;68(20):8333-41]

    DOI: 10.1158/0008-5472.CAN-08-0080

    Web of Science

    researchmap

  • The structure of S100A11 fragment explains a local structural change induced by phosphorylation

    Takahide Kouno, Mineyuki Mizuguchi, Masakiyo Sakaguchi, Eiichi Makino, Yoshihiro Mori, Hiroyuki Shinoda, Tomoyasu Aizawa, Makoto Demur, Nam-Ho Huh, Keiichi Kawano

    JOURNAL OF PEPTIDE SCIENCE   14 ( 10 )   1129 - 1138   2008.10

     More details

    Language:English   Publisher:JOHN WILEY & SONS LTD  

    S100A11 protein is a member of the S100 family containing two EF-hand motifs. It undergoes phophorylation on residue T10 after cell stimulation such as an increase in Ca2+ concentration. Phosphorylated S100A11 I can be recognized by its target protein, nucleolin. Although S100A11 is initially expressed in the cytoplasm, it is transported to the nucleus by the action of nucleolin. In the nucleus, S100A11 suppresses the growth of keratinocytes through p21(CIP1/WAF1) activation and induces cell differentiation. Interestingly, the N-terminal fragment of S100A11 has the same activity as the full-length protein: i.e. it is phosphorylated in vivo and binds to nucleolin. In addition, this fragment leads to the arrest of cultured keratinocyte growth. We examined the solution structure of this fragment peptide and explored its structural properties before and after phosphorylation. In a trifluoroethanol solution, the peptide adopts the alpha-helical structure just as the corresponding region of the full-length S100A11. Phosphorylation induces a disruption of the N-capping conformation of the alpha-helix, and has a tendency to perturb its surrounding structure. Therefore, the phosphorylated threonine lies in the N-terminal edge of the alpha-helix. This local structural change can reasonably explain why the phosphorylation of a residue that is initially buried in the interior of protein allows it to be recognized by the binding partner. Copyright (C) 2008 European Peptide Society and John Wiley & Sons, Ltd.

    DOI: 10.1002/psc.1050

    Web of Science

    researchmap

  • Intracellular delivery of glutathione S-transferase-fused proteins into mammalian cells by polyethylenimine-glutathione conjugates

    Hitoshi Murata, Junichiro Futami, Midori Kitazoe, Takayuki Yonehara, Hidetaka Nakanishi, Megumi Kosaka, Hiroko Tada, Masakiyo Sakaguchi, Yasuyuki Yagi, Masaharu Seno, Nam-ho Huh, Hidenori Yamada

    JOURNAL OF BIOCHEMISTRY   144 ( 4 )   447 - 455   2008.10

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    The glutathione S-transferase (GST)-fused protein expression system has been extensively used to generate a large quantity of proteins and has served for functional analysis in vitro. In this study, we developed a novel approach for the efficient intracellular delivery of GST-fused proteins into living cells to expand their usefulness up to in vivo use. Since protein cationization techniques are powerful strategies for efficient intracellular uptake by adsorptive-mediated endocytosis, GST-fused proteins were cationized by forming a complex with a polycationic polyethylenimine (PEI)-glutathione conjugate. On screening of protein transduction, optimized PEI-glutathione conjugate for protein transduction was characterized by a partly oligomerized mixture of PEI with average molecular masses of 600 (PEI600) modified with multiple glutathiones, which could have sufficient avidity for GST. Furthermore, enhanced endosomal escape of transduced GST-fused proteins was observed when they were delivered with a glutathione-conjugated PEI600 derivative possessing a hydroxybutenyl moiety. These results were confirmed by both intracellular confocal imaging of GST-fused green fluorescent protein and activation of an endogenous growth signal transduction pathway by a GST-fused constitutively active mutant of a kinase protein. These PEI-glutathione conjugates seem to be convenient molecular tools for protein transduction of widely used GST-fused proteins.

    DOI: 10.1093/jb/mvn087

    Web of Science

    researchmap

  • Derivation of hepato-pancreatic intermediate progenitor cells from a clonal mesenchymal stem cell line of rat bone marrow origin

    Takuro Masaka, Masahiro Miyazaki, Gang Du, Marhaen Hardjo, Masakiyo Sakaguchi, Mikiro Takaishi, Ken Kataoka, Kazuhide Yamamoto, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   22 ( 4 )   447 - 452   2008.10

     More details

    Language:English   Publisher:PROFESSOR D A SPANDIDOS  

    We have recently established a clonal mesenchymal stem cell line (rBM25/S3) from adult rat bone marrow. The cells have practically unlimited proliferation capacity (over 300 PDL), maintaining multipotency for differentiation. In the present study, we examined the potential for rBM25/S3 cells to differentiate into insulin-secreting cells. When cultured in the presence of HGF and FGF-4 on Matrigel, rBM25/S3 cells expressed genes specific to pancreatic B-cells as well as those specific to hepatocytes. They still maintained proliferation capacity with a doubling time of similar to 30 h. These hepato-pancreatic intermediate progenitor cells, but not the original undifferentiated rBM25/S3 cells, were induced by the overexpression of PDX-1 to produce significant amounts of insulin in a manner responding to glucose concentration in medium. The present culture system indicates a direction for further studies aimed at the realization of cell transplantation therapy for type I diabetes mellitus.

    DOI: 10.3892/ijmm_00000041

    Web of Science

    researchmap

  • An N-terminal 78 amino acid truncation of REIC/Dkk-3 effectively induces apoptosis

    Fernando Abarzua, Yuji Kashiwakura, Munenori Takaoka, Masami Watanabe, Kazuhiko Ochiai, Masakiyo Sakaguchi, Takao Iwawaki, Ryuta Tanimoto, Yasutomo Nasu, Nam-ho Huh, Hiromi Kumon

    BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS   375 ( 4 )   614 - 618   2008.10

     More details

    Language:English   Publisher:ACADEMIC PRESS INC ELSEVIER SCIENCE  

    Overexpression of REIC/Dkk-3 (a tumor suppressor gene) induces cancer cell apoptosis through endoplasmic reticulum (ER) stress. Therefore, the identification of the portion of REIC/Dkk-3 that causes ER stress may be essential for the development of cancer treatment based on REIC/Dkk-3. Here, we made several truncated forms of REIC/Dkk-3 and investigated their therapeutic potentials against prostate cancer. Among three truncated forms, a variant comprising the N-terminal 78 amino acid region of REIC/Dkk-3 ((1-78) REIC/Dkk-3) most strongly induced ER stress and apoptosis in human prostate cancer cells (PC3). For in vivo gene expression, we coupled a biodegradable polymer with naked DNA, which attained robust trans-gene expression in PC3-derived subcutaneous tumor. In therapeutic experiments, we demonstrated that multiple direct injections of polymer-conjugated (1-78)REIC/Dkk-3 plasmid provoke ER stress and significantly reduced the subcutaneous tumor volume compared with the control group. We suggest this non-viral strategy may be an effective alternative to viral gene therapy. (C) 2008 Elsevier Inc. All rights reserved.

    DOI: 10.1016/j.bbrc.2008.08.079

    Web of Science

    researchmap

  • REIC/Dkk-3遺伝子治療の正常細胞を介する抗腫瘍効果(Transduction of gene therapeutic REIC/Dkk-3 into normal cells provides another arm for tumor suppression in vivo)

    片岡 健, 阪口 政清, Fernando Abarzua, 谷本 竜太, 渡部 昌実, 村田 等, 那須 保友, 公文 裕巳, 許 南浩

    日本癌学会総会記事   67回   93 - 93   2008.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • REIC/Dkk-3遺伝子によるアポトーシス耐性メカニズムとその克服(BiP is a determining protein for sensitivity to apoptosis induced by a gene-therapeutic adenovirus carrying REIC/Dkk-3)

    谷本 竜太, 阪口 政清, アバルズア・フェルナンド, 片岡 健, 那須 保友, 公文 裕巳, 許 南浩

    日本癌学会総会記事   67回   238 - 238   2008.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • REIC/Dkk-3の強制発現はERストレスを介してIL-7を誘導する(Over-expression of REIC/Dkk-3 induces IL-7 in normal human fibroblasts via ER stress)

    阪口 政清, 片岡 健, アバルスア・フェルナンド, 谷本 竜太, 渡辺 昌実, 村田 等, 那須 保友, 公文 裕己, 許 南浩

    日本癌学会総会記事   67回   238 - 238   2008.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • BRPK/PINK1はAktのリン酸化によってがん細胞のアポトーシスを抑制する(BRPK/PINK1 blocks apoptotic cell death of cancer cells by phosphotylation of Akt)

    村田 等, 阪口 政清, 片岡 健, 許 南浩

    日本癌学会総会記事   67回   64 - 64   2008.9

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • S100A8/A9, a key mediator for positive feedback growth stimulation of normal human keratinocytes

    Takamasa Nukui, Ritsuko Ehama, Masakiyo Sakaguchi, Hiroyuki Sonegawa, Chika Katagiri, Toshihiko Hibino, Nam-Ho Huh

    JOURNAL OF CELLULAR BIOCHEMISTRY   104 ( 2 )   453 - 464   2008.5

     More details

    Language:English   Publisher:WILEY-LISS  

    S100A8 and S100A9 are known to be up-regulated in hyperproliferative and psoriatic epidermis, but their function in epidermal keratinocytes remains largely unknown. Here we show that (1) S100A8 and S100A9 are secreted by cultured normal human keratinocytes (NHK) in a cytokine-dependent manner, (2) when applied to NHK, recombinant S100A8/A9 (a 1:1 mixture of S100A8 and S100A9) induced expression of a number of cytokine genes such as IL-8/CXCL8, CXCL1, CXCL2, CXCL3, CCL20, IL-6, and TNF alpha that are known to be up-regulated in psoriatic epidermis, (3)the S100A8/ A9-induced cytokines in turn enhanced production and secretion of S100A8 and S100A9 by NHK, and (4) S100A8 and S100A8/A9 stimulated the growth of NHK at a concentration as low as 1 ng/ml. These results indicate the presence of a positive feedback loop for growth stimulation involving S100A8/A9 and cytokines in human epidermal keratinocytes, implicating the relevance of the positive feedback loop to the etiology of hyperproliferative skin diseases, including psoriasis.

    DOI: 10.1002/jcb.21639

    Web of Science

    researchmap

  • S100A11 functions as a dual mediator for growth regulation in normal human keratinocytes

    M. Sakaguchi, H. Murata, N. Huh

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   128   S138 - S138   2008.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:NATURE PUBLISHING GROUP  

    Web of Science

    researchmap

  • S100A11, an dual mediator for growth regulation of human keratinocytes

    Masakiyo Sakaguchi, Hiroyuki Sonegawa, Hitoshi Murata, Midori Kitazoe, Jun-ichiro Futami, Ken Kataoka, Hidenori Yamada, Nam-ho Huh

    MOLECULAR BIOLOGY OF THE CELL   19 ( 1 )   78 - 85   2008.1

     More details

    Language:English   Publisher:AMER SOC CELL BIOLOGY  

    We previously revealed a novel signal pathway involving S100A11 for inhibition of the growth of normal human keratinocytes (NHK) caused by high Ca++ or transforming growth factor beta. Exposure to either agent resulted in transfer of S100A11 to nuclei, where it induced p21(WAF1). In contrast, S100A11 has been shown to be overexpressed in many human cancers. To address this apparent discrepancy, we analyzed possible new functions of S100A11, and we provide herein evidence that 1) S100A11 is actively secreted by NHK; 2) extracellular S100A11 acts on NHK to enhance the production of epidermal growth factor family proteins, resulting in growth stimulation; 3) receptor for advanced glycation end products, nuclear factor-kappa B, Akt, and cAMP response element-binding protein are involved in the S100A11-triggered signal transduction; and 4) production and secretion of S100A11 are markedly enhanced in human squamous cancer cells. These findings indicate that S100A11 plays a dual role in growth regulation of epithelial cells.

    DOI: 10.1091/mbc.E07-07-0682

    Web of Science

    researchmap

  • Truncation of annexin A1 is a regulatory lever for linking epidermal growth factor signaling with cytosolic phospholipase A2 in normal and malignant squamous epithelial cells

    Masakiyo Sakaguchi, Hitoshi Murata, Hiroyuki Sonegawa, Yoshihiko Sakaguchi, Jun-ichiro Futami, Midori Kitazoe, Hidenori Yamada, Nam-ho Huh

    JOURNAL OF BIOLOGICAL CHEMISTRY   282 ( 49 )   35679 - 35686   2007.12

     More details

    Language:English   Publisher:AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC  

    Regulation of cell growth and apoptosis is one of the pleiotropic functions of annexin A1 (ANXA1). Although previous reports on the overexpression of ANXA1 in many human cancers and on growth suppression and/or induction of apoptosis by ANXA1 may indicate the tumor-suppressive nature of ANXA1, molecular mechanisms of the function of ANXA1 remain largely unknown. Here we provide evidence that ANXA1 mechanistically links the epidermal growth factor-triggered growth signal pathway with cytosolic phospholipase A(2) (cPLA(2)), an initiator enzyme of the arachidonic acid cascade, through interaction with S100A11 in normal human keratinocytes (NHK). Ca2+ -dependent binding of S100A11 to ANXA1 facilitated the binding of the latter to cPLA2, resulting in inhibition of cPLA2 activity, which is essential for the growth of NHK. On exposure of NHK to epidermal growth factor, ANXA1 was cleaved solely at Trp(12), and this cleavage was executed by cathepsin D. In squamous cancer cells, this pathway was shown to be constitutively activated. The newly found mechanistic intersection may be a promising target for establishing new measures against human cancer and other cell growth disorders.

    DOI: 10.1074/jbc.M707538200

    Web of Science

    researchmap

  • Isolation of a bone marrow-derived stem cell line with high proliferation potential and its application for preventing acute fatal liver failure

    Masahiro Miyazaki, Marhaen Hardjo, Takuro Masaka, Koji Tomiyama, Naila Mahmut, Reinhold J. Medina, Aya Niida, Hiroyuki Sonegawa, Gang Du, Rong Yong, Mikiro Takaishi, Masakiyo Sakaguchi, Nam-Ho Huh

    STEM CELLS   25 ( 11 )   2855 - 2863   2007.11

     More details

    Language:English   Publisher:ALPHAMED PRESS  

    Transplantation of hepatocytes or hepatocyte-like cells of extrahepatic origin is a promising strategy for treatment of acute and chronic liver failure. We examined possible utility of hepatocyte-like cells induced from bone marrow cells for such a purpose. Clonal cell lines were established from the bone marrow of two different rat strains. One of these cell lines, rBM25/S3 cells, grew rapidly (doubling time, similar to 24 hours) without any appreciable changes in cell properties for at least 300 population doubling levels over a period of 300 days, keeping normal diploid karyotype. The cells expressed CD29, CD44, CD49b, CD90, vimentin, and fibronectin but not CD45, indicating that they are of mesenchymal cell origin. When plated on Matrigel with hepatocyte growth factor and fibroblast growth factor-4, the cells efficiently differentiated into hepatocyte-like cells that expressed albumin, cytochrome P450 (CYP) 1A1, CYP1A2, glucose 6-phosphatase, tryptophane-2,3-dioxygenase, tyrosine aminotransferase, hepatocyte nuclear factor (HNF)1 alpha, and HNF4 alpha. Intrasplenic transplantation of the differentiated cells prevented fatal liver failure in 90%-hepatectomized rats. In conclusion, a clonal stem cell line derived from adult rat bone marrow could differentiate into hepatocyte-like cells, and transplantation of the differentiated cells could prevent fatal liver failure in 90%-hepatectomized rats. The present results indicate a promising strategy for treating human fatal liver diseases.

    DOI: 10.1634/stemcells.2007-0078

    Web of Science

    researchmap

  • ヒト膀胱癌細胞におけるREIC/Dkk-3誘導性アポトーシスに対する抵抗性の機構解析(Mechanistic Analysis of Resistance to REIC/Dkk-3-induced Apoptosis in Human Bladder Cancer Cell Lines)

    小林 知子, 阪口 政清, 谷本 竜太, 枝村 康平, アバラズア・フェルナンド, 片岡 健, 賀来 春紀, 雑賀 隆史, 宮崎 正博, 那須 保友, 公文 裕巳, 許 南浩

    西日本泌尿器科   69 ( 増刊 )   125 - 125   2007.10

     More details

    Language:English   Publisher:西日本泌尿器科学会  

    researchmap

  • Adenovirus-mediated REIC/Dkk-3 gene transfer inhibits tumor growth and metastasis in an orthotopic prostate cancer model

    K. Edamura, Y. Nasu, M. Takaishi, T. Kobayashi, F. Abarzua, M. Sakaguchi, Y. Kashiwakura, S. Ebara, T. Saika, M. Watanabe, N-H Huh, H. Kumon

    CANCER GENE THERAPY   14 ( 9 )   765 - 772   2007.9

     More details

    Language:English   Publisher:NATURE PUBLISHING GROUP  

    We had previously reported that REIC/Dkk-3, a member of the Dickkopf ( Dkk) gene family, works as a tumor suppressor. In this study, we evaluated the therapeutic effects of an intratumoral injection with adenoviral vector encoding REIC/Dkk-3 gene ( Ad-REIC) using an orthotopic mouse prostate cancer model of RM-9 cells. We also investigated the in vivo anti-metastatic effect and in vitro anti-invasion effect of Ad-REIC gene delivery. We demonstrated that the Ad-REIC treatment inhibited prostate cancer growth and lymph node metastasis, and prolonged mice survival in the model. These therapeutic responses were consistent with the intratumoral apoptosis induction and in vitro suppression of cell invasion/migration with reduced matrix metalloprotease-2 activity. We thus concluded that in situ Ad-REIC/Dkk-3 gene transfer may be a promising therapeutic intervention modality for the treatment of prostate cancer.

    DOI: 10.1038/sj.cgt.7701071

    Web of Science

    researchmap

  • ヒト膀胱癌細胞株におけるREIC/Dkk-3誘導性アポトーシスに対する抵抗性の機構的解析(Mechanistic Analysis of Resistance to REIC/Dkk-3-induced Apoptosis in Human Bladder Cancer Cell Lines)

    小林 知子, 阪口 政清, 谷本 竜太, アバラズア・フェルナンド, 高石 樹朗, 片岡 健, 賀来 春紀, 雑賀 隆史, 那須 保友, 宮崎 正博, 公文 裕巳, 許 南浩

    日本癌学会総会記事   66回   476 - 476   2007.8

     More details

    Language:English   Publisher:日本癌学会  

    researchmap

  • Heat shock proteins play a crucial role in tumor-specific apoptosis by REIC/Dkk-3

    Fernando Abarzua, Masakiyo Sakaguchi, Ryuta Tanimoto, Hiroyuki Sonegawa, Dai-Wei L, Kohei Edamura, Tomoko Kobayashi, Masami Watanabe, Yuji Kashiwakura, Haruki Kaku, Takashi Saika, Keiichiro Nakamura, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   20 ( 1 )   37 - 43   2007.7

     More details

    Language:English   Publisher:PROFESSOR D A SPANDIDOS  

    We recently showed that overexpression of REIC/Dickkopf-3 (Dkk-3), a tumor suppressor gene, induced apoptosis in a tumor cell-specific manner. The aim of the present study was to determine the mechanisms underlying the selective induction of apoptosis. At first, we found a mouse renal carcinoma cell line, RENCA, to be extremely sensitive to an adenovirus carrying REIC/Dkk-3 (Ad-REIC), and we showed that activation of c-Jun N-terminal kinase (JNK) was a critical step in cell death, i.e. a process similar to that in human prostate and testicular cancer observed in our previous studies. Among the proteins interfering with the activation of JNK, heat shock protein (Hsp)70/72 was reduced in expression in RENCA cells compared with that in NIH3T3 cells. An Hsp70/72 inducer protected RENCA cells from Ad-REIC-induced apoptosis, while an Hsp70/72 inhibitor sensitized NIH3T3 cells for apoptosis induction. These results indicate that functionally active Hsp70/72 is a key factor in tumor cell-specific induction of apoptotic cell death and that analyses of the expression levels of Hsp70/72 may be essential in determining the significance of Ad-REIC-based gene therapy against human cancer.

    Web of Science

    researchmap

  • Involvement of deterioration in S100C/A11-mediated pathway in resistance of human squamous cancer cell lines to TGF beta-induced growth suppression

    Hiroyuki Sonegawa, Takamasa Nukui, Dai-Wei Li, Mikiro Takaishi, Masakiyo Sakaguchi, Nam-Ho Hub

    JOURNAL OF MOLECULAR MEDICINE-JMM   85 ( 7 )   753 - 762   2007.7

     More details

    Language:English   Publisher:SPRINGER  

    Recently, we demonstrated that S100C/A11 comprises an essential pathway for growth suppression by TGF beta in normal human keratinocytes. Nuclear transfer of S 100C/A11 was a hallmark of the activation of the process. In the present study, we examined the possible deterioration in the pathway in human squamous cancer cell lines, focusing on intracellular localization of S100C/A11 and its functional partners Smad3 and Smad4. All four human squamous cancer cell lines examined (A431, BSCC-93, DJM-1, and HSC-5) were resistant to growth suppression by TGF beta. In BSCC-93, DJM-1, and HSC-5 cells exposed to TGF beta, S100C/A11 was not transferred to the nuclei, and p21(WAF1) was not induced. Overexpression of nucleus-targeted S100C/A11 partially recovered induction of p21 (WAF1) and p15(INK4B) and growth suppression by TGF beta 1 in these cells. These results indicate that the deterioration in the S100C/A11-mediated pathway conferred upon the cancer cell lines resistance to TGF beta. In A431 cells, S100C/A11, Smad3, and Smad4 were simultaneously transferred to the nuclei, and p2l(WAF1) was induced upon exposure to TGF beta. We provide evidence to indicate that refractoriness of A431 cells to TGF beta was probably because the amount of p2l(WAF1) induced by TGF beta was insufficient to counteract cyclin A, which is highly overexpressed in A431 cells. Thus, the newly found S100C/A11-mediated pathway is at least partly involved in conferring upon human squamous cell cancers resistant to TGF beta-induced growth suppression, which is considered to play a critical role for the initiation and progression of many human cancers.

    DOI: 10.1007/s00109-007-0180-7

    Web of Science

    researchmap

  • S100A8/A9, a key mediator for positive feedback growth stimulation and inflammation of normal human keratinocytes

    R. Ehama, T. Nukui, C. Katagiri, M. Sakaguchi, H. Sonegawa, N. Huh, T. Hibino

    JOURNAL OF INVESTIGATIVE DERMATOLOGY   127   S98 - S98   2007.4

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:NATURE PUBLISHING GROUP  

    Web of Science

    researchmap

  • REIC/Dkk-3 as a potential gene therapeutic agent against human testicular cancer

    Ryuta Tanimoto, Fernando Abarzua, Masakiyo Sakaguchi, Mikiro Takaishi, Yasutomo Nasu, Hiromi Kumon, Nam-Ho Huh

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   19 ( 3 )   363 - 368   2007.3

     More details

    Language:English   Publisher:PROFESSOR D A SPANDIDOS  

    Human testicular cancer is very sensitive to chemotherapy and radiation therapy and is regarded as a curable cancer. The cancer prevails in the young reproductive generation and testicular dysfunction is often observed as a side effect, remaining a serious challenge. In the present study, we examined the potential utility of REIC/Dkk-3-based gene therapy against human testicular cancer. Expression of REIC/Dkk-3 was reduced in all of the human seminoma and non-seminomatous germ cell tumor tissues. Overexpression of REIC/Dkk-3 using an adenovirus vector (Ad-REIC) induced apoptosis in a testicular germ cell cancer cell line NCCIT but not in normal human fibroblasts. c-Jun terminal kinase (JNK) was activated by Ad-REIC and the induction of apoptosis was abrogated by a JNK inhibitor. A single intratumoral injection of Ad-REIC markedly inhibited the tumorigenic growth of NCCIT cells in nude mice. These results indicate that Ad-REIC may lead to developing less insulting and non-genotoxic therapeutic measures against human testicular cancer.

    Web of Science

    researchmap

  • Denatured and reversibly cationized p53 readily enters cells and simultaneously folds to the functional protein in the cells

    H Murata, M Sakaguchi, J Futami, M Kitazoe, T Maeda, H Doura, M Kosaka, H Tada, M Seno, N Huh, H Yamada

    BIOCHEMISTRY   45 ( 19 )   6124 - 6132   2006.5

     More details

    Language:English   Publisher:AMER CHEMICAL SOC  

    Cationization is a powerful strategy for internalizing a protein into living cells. On the other hand, a reversibly cationized denatured protein through disulfide bonds is not only soluble in water but also able to fold to the native conformation in vitro. When these advantages in cationization were combined, we developed a novel method to deliver a denatured protein into cells and simultaneously let it fold to express its function within cells. This "in-cell folding" method enhances the utility of recombinant proteins expressed in Escherichia coli as inclusion bodies; that is, the recombinant proteins in inclusion bodies are solubilized by reversible cationization through cysteine residues by disulfide bonds with aminopropyl methanethiosulfonate or pyridyldithiopropionylpolyethylenimine and then incubated with cells without an in vitro folding procedure. As a model protein, we investigated human tumor-suppressor p53. Treatment of p53-null Saos-2 cells with reversibly cationized p53 revealed that all events examined as indications of the activation of p53 in cells, such as reduction of disulfide bonds followed by tetramer formation, localization into the nucleus, induction of p53 target genes, and induction of apoptosis of cells, occurred. These results suggest that reversible cationization of a denatured protein through cysteine residues is an alternative method for delivery of a functional protein into cells. This method would be very useful when a native folded protein is not readily available.

    DOI: 10.1021/bi052642a

    Web of Science

    researchmap

  • HNF4 転写因子による肝機能制御機構の解明

    阪口政清

    ファルマシア   2006

     More details

  • Adenovirus-mediated overexpression of REIC/Dkk-3 selectively induces apoptosis in human prostate cancer cells through activation of c-Jun-NH2-kinase

    F Abarzua, M Sakaguchi, M Takaishi, Y Nasu, K Kurose, S Ebara, M Miyazaki, M Namba, H Kumon, N Huh

    CANCER RESEARCH   65 ( 21 )   9617 - 9622   2005.11

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    Alteration in genes which takes place during malignant conversion and progression could be potential targets for gene therapy. We previously identified REIC/Dkk-3 as a gene whose expression is reduced in many human cancers. Here, we showed that expression of REIC/Dkk-3 was consistently reduced in human prostate cancer tissues in a stage-dependent manner. Forced expression of REIC/Dkk-3 induced apoptosis in human prostate cancer cell lines lacking endogenous REIC/Dkk-3 expression but not in REIC/Dkk3-proficient normal prostate epithelial and stromal cells. The apoptosis involved c-jun-NH2-kinase activation, mitochondrial translocation of Bax, and reduction of Bcl-2. A single injection of an adenovirus vector carrying REIC/Dkk-3 showed a dramatic antitumor effect on a xenotransplanted human prostate cancer. Thus, REIC/Dkk-3 could be a novel target for gene-based therapy of prostate cancer.

    DOI: 10.1158/0008-5472.CAN-05-0829

    Web of Science

    researchmap

  • Bifurcated converging pathways for high Ca2+- and TGF beta-induced inhibition of growth of normal human keratinocytes

    M Sakaguchi, H Sonegawa, T Nukui, Y Sakaguchi, M Miyazaki, M Namba, N Huh

    PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA   102 ( 39 )   13921 - 13926   2005.9

     More details

    Language:English   Publisher:NATL ACAD SCIENCES  

    Growth suppression of normal human keratinocytes by high Ca2+ or TGF beta was shown to be mediated by p2l(WAF1/C1P1) and Sp1 [Pardali, K., et al. (2000) J. Biol. Chem. 275, 29244-29256; Santini, M. P., Talora, C., Seki, T., Bolgan, L. & Dotto, G. P. (2001) Proc. Nat. Acad Sci. USA 98, 9575-9580; Al-Daraji, W. I., Grant, K. R., Ryan, K., Saxton, A., & Reynolds, N. J. (2002) J. Invest. Dermato/. 118, 779-788]. We previously demonstrated that S100C/A11 is a key mediator for growth inhibition of normal human epidermal keratinocytes (NHK) triggered by high Ca2+ or TGF beta [Sakaguchi, M., et al. (2003) J. Cell Biol. 163, 825-835; Sakaguchi, M., et al. (2004) 164, 979-984]. On exposure of NHK cells to either agent, S100C/ All is transferred to nuclei, where it induces p21(WAF1/CIP1) through activation of Sp1/Sp3. In the present study, we found that high Ca2+ activated NFAT1 through calcineurin-dependent dephosphorylation. In growing NHK cells, Krueppel-like factor (KLF)16, a member of the Sp/KLF family, bound to the p21(WAF1/CIP1) promoter and, thereby, inhibited the transcription of p21(WAF1/CIP1). Sp1 complexed with NFAT1 in high Ca2+ -treated cells or with Smad3 in TGF beta 1-treated cells, but not Sp1 alone, replaced KLF16 from the p21(WAF1/CIP1) promoter and transcriptionally activated the p21(WAF1/CIP1) gene. Thus, high Ca2+ and TGF beta 1 have a common S1OOC/A11-mediated pathway in addition to a unique pathway (NFAT1-mediated pathway for high Ca2+ and Smad-mediated pathway for TGF beta 1) for exhibiting a growth inhibitory effect on NHK cells, and both pathways were shown to be indispensable for growth inhibition.

    DOI: 10.1073/pnas.0500630102

    Web of Science

    researchmap

  • Protein transduction assisted by polyethylenimine-cationized carrier proteins

    M Kitazoe, H Murata, J Futami, T Maeda, M Sakaguchi, M Miyazaki, M Kosaka, H Tada, M Seno, N Huh, M Namba, M Nishikawa, Y Maeda, H Yamada

    JOURNAL OF BIOCHEMISTRY   137 ( 6 )   693 - 701   2005.6

     More details

    Language:English   Publisher:JAPANESE BIOCHEMICAL SOC  

    Previously, we have reported that cationized-proteins covalently modified with polyethylenimine (PEI) (direct PEI-cationization) efficiently enter cells and function in the cytosol [Futami et al. (2005) J. Biosci. Bioeng. 99,95-1031. However, it may be more convenient if a protein could be delivered into cells just by mixing the protein with a PEI-cationized carrier protein having a specific affinity (indirect PEI-cationization). Thus, we prepared PEI-cationized avidin (PEI-avidin), streptavidin (PEI-streptavidin), and protein G (PEI-protein G), and examined whether they could deliver biotinylated proteins and antibodies into living cells. PEI-avidin (and/or PEI-streptavidin) carried biotinylated GFPs into various mammalian cells very efficiently. A GFP variant containing a nuclear localization signal was found to arrive even in the nucleus. The addition of a biotinylated RNase A derivative mixed with PEI-streptavidin to a culture medium of 3T3-SV-40 cells resulted in remarkable cell growth inhibition, suggesting that the biotinylated RNase A derivative entered cells and digested intracellular RNA molecules. Furthermore, the addition of a fluorescein-labeled antiS100C (beta-actin binding protein) antibody mixed with PEI-protein G to human fibroblasts resulted in the appearance of a fluorescence image of actin-like filamentous structures in the cells. These results indicate that indirect PEI-cationization using non-covalent interaction is as effective as the direct PEI-cationization for the transduction of proteins into living cells and for expression of their functions in the cytosol. Thus, PEI-cationized proteins having a specific affinity for certain molecules such as PEI-streptavidin, PEI-avidin and PEI-protein G are concluded to be widely applicable protein transduction carrier molecules.

    DOI: 10.1093/jb/mvi081

    Web of Science

    researchmap

  • Intracellular delivery of proteins into mammalian living cells by polyethylenimine-cationization

    J Futami, M Kitazoe, T Maeda, E Nukui, M Sakaguchi, J Kosaka, M Miyazaki, M Kosaka, H Tada, M Seno, Y Sasaki, NH Huh, M Namba, H Yamada

    JOURNAL OF BIOSCIENCE AND BIOENGINEERING   99 ( 2 )   95 - 103   2005.2

     More details

    Language:English   Publisher:SOC BIOSCIENCE BIOENGINEERING JAPAN  

    In the post-genomic era, there is pressing need for development of protein manipulation methodology to analyze functions of proteins in living cells. For this purpose, techniques to deliver functional proteins into living cells are currently being evaluated as alternative approaches to the introduction of transcriptionally active DNA. Here, we describe a novel method for efficient protein transduction into living cells in which a protein is simply cationized with polyethylenimine (PEI) by limited chemical conjugation. PEI-cationized proteins appear to adhere to the cell surface by ionic charge interaction and then internalize into cells in a receptor- and transporter-independent fashion. Since PEI is an organic macromolecule with a high cationic-charge density, limited coupling with PEI results in endowment of sufficient cationic charge to proteins without causing serious decline in their fundamental functions. A number of PEI-cationized proteins, such as ribonuclease (RNase), green fluorescent protein (GFP) and immunoglobulin (IgG), efficiently entered cells and functioned in the cytosol. Our results suggest that protein cationization techniques using PEI will be useful for the development of protein transduction technology.

    DOI: 10.1263/jbb.99.095

    Web of Science

    researchmap

  • Targeted disruption of transcriptional regulatory function of p53 by a novel efficient method for introducing a decoy oligonucleotide into nuclei

    M Sakaguchi, T Nukui, H Sonegawa, H Murata, J Futami, H Yamada, N Huh

    NUCLEIC ACIDS RESEARCH   33 ( 9 )   2005

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    Decoy oligonucleotides have been used for functional sequestering of transcription factors. Efficient introduction into cells is a prerequisite for the oligonucleotides to exert their blocking function. Lipofection is the most widely used technique for that purpose because of its convenience and relatively high efficiency. However, the transduction efficiency of lipofection largely depends on cell types and experimental conditions and the introduced nucleotides are not specifically directed to nuclei where they exert their major function. In the present study, we designed a new system for transporting oligonucleotides into cell nuclei. The vehicle is composed of glutathione-S-transferase, 7 arginine residues, the DNA- binding domain of GAL4 and a nuclear localization signal, which are linked with flexible glycine stretches. The p53-responsive element linked to the GAL4 upstream activating sequence was efficiently transferred by the vehicle protein into nuclei of primary cultures of neuronal cells, embryonic stem cells and various human normal cells. Transcriptional activation of p21(WAF1/CIP1) and Bax by p53 on exposure to cisplatin was completely blocked by introducing the p53 decoy oligonucleotide. Thus, the system developed in the present study can be a convenient and powerful tool for specifically disrupting the function of DNA- binding proteins in culture.

    DOI: 10.1093/nar/gni088

    Web of Science

    researchmap

  • siRNAによる遺伝子発現抑制法

    阪口政清, 曽根川裕之, 温井孝昌, 許 南浩

    Surgery Frontier   2005

     More details

  • PKCa mediates TGFb-induced growth inhibition of human keratinocytes via phosphorylation of S100C/A11

    M Sakaguchi, M Miyazaki, H Sonegawa, M Kashiwagi, M Ohba, T Kuroki, M Namba, N Huh

    MOLECULAR BIOLOGY OF THE CELL   15   449A - 449A   2004.11

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:AMER SOC CELL BIOLOGY  

    Web of Science

    researchmap

  • Increased expression of calcium-binding protein S100 in human uterine smooth muscle tumours

    T Kanamori, K Takakura, M Mandai, M Kariya, K Fukuhara, M Sakaguchi, NH Huh, K Saito, T Sakurai, J Fujita, S Fujii

    MOLECULAR HUMAN REPRODUCTION   10 ( 10 )   735 - 742   2004.10

     More details

    Language:English   Publisher:OXFORD UNIV PRESS  

    S100 proteins belong to the EF-hand Ca2+ -binding protein family and regulate a variety of cellular processes via interaction with different target proteins. Several diseases, including cancer and melanoma, are related to the abnormal expression of S100 proteins, which are expressed in cell- and tissue-specific manners. We investigated the expression of S100 family members in human uterine smooth muscle tumours. Expression of six members of the S100 protein family: S100A1, A4, A6, A7, A10 and A11, was found in human uterine leiomyoma and myometrium tissue, but expression of other members was not detected by RT-PCR. Real-time PCR showed that S100A11 expression was significantly increased in leiomyoma compared with myometrium. Suppression of S100A11 by small interfering RNA (siRNA) led to apoptosis, and the overexpression of S100A11 inhibited apoptosis in human uterine smooth muscle tumour cells. These findings suggest that S100A11 has an anti-apoptotic function and is related to the process of growth of human uterine leiomyoma.

    DOI: 10.1093/molehr/gah100

    Web of Science

    researchmap

  • Expression of CYP3A4 by an immortalized human hepatocyte line in a three-dimensional culture using a radial-flow bioreactor

    Akiyama, I, K Tomiyama, M Sakaguchi, M Takaishi, M Mori, M Hosokawa, S Nagamori, N Shimizu, NH Huh, M Miyazaki

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   14 ( 4 )   663 - 668   2004.10

     More details

    Language:English   Publisher:PROFESSOR D A SPANDIDOS  

    Cytochrome P450 (CYP) 3A is responsible for about 50% of drug metabolizing activity in the liver. The present study was undertaken to establish a CYP3A4-active model for in vitro analysis of human drug metabolism. The cells used were immortalized normal human fetal hepatocytes (OUMS-29) and its HNF4alpha-introduced subline (OUMS-29/H-11). The cells were cultivated under high-density three-dimensional conditions in a radial-flow bioreactor (RFB). The number of OUMS-29 cells increased 15-fold over 49 days and their apical surfaces were covered with abundant microvilli, a characteristic of hepatocytes in vivo. The amount of albumin secreted by OUMS-29 cells in the three-dimensional RFB culture was 6-fold higher than those in a monolayer culture. CYP3A4 protein and an intermediate metabolite of testosterone by CYP3A4 were detected in OUMS-29/H11 cells cultivated in RFB &gt;29 days. These results indicate that the RFB; culture of OUMS-29/H-11 cells is useful for screening and developing new drugs.

    Web of Science

    researchmap

  • Introduction of an N-terminal peptide of S100C/A11 into human cells induces apoptotic cell death

    E Makino, M Sakaguchi, K Iwatsuki, N Huh

    JOURNAL OF MOLECULAR MEDICINE-JMM   82 ( 9 )   612 - 620   2004.9

     More details

    Language:English   Publisher:SPRINGER  

    S100 proteins belong to the EF-hand Ca2+- binding protein family and are involved in the regulation of a variety of cellular processes. Individual S 100 proteins are expressed in cell- and tissue-specific manners, and functional deterioration of S100 proteins leads to a number of human diseases, including cancer. We previously demonstrated that S100C/A11 was translocated to nuclei and inhibited DNA synthesis in human keratinocytes when exposed to high Ca2+. In the present study we examined the effects of synthetic partial peptides of S100C/ A11 on human carcinoma cell lines. Only an N-terminal peptide with 19 amino acid residues (MAK19) showed cytotoxicity to the cell lines in dose- and time-dependent manners when introduced into cells by flanking the HIV-TAT protein transduction domain (TAT-MAK19). Pulse field electrophoresis revealed that DNA of the treated cells was partially degradated. Annexin V, a marker of cellular apoptosis, was detected in the cells treated with TAT-MAK19 by immunostaining and flow cytometry. The induction of apoptotic cell death was apparently independent of p53, p21(WAF1/CIP1), and caspase activity, but treatment with TAT-MAK19 resulted in partial translocation of apoptosis-inducing factor (AIF) from the cytoplasm to nuclei. These results indicate that MAK19 induces apoptosis in human cell lines and may therefore lead to the establishment of a new molecular target for the treatment of human cancer.

    DOI: 10.1007/s00109-004-0560-1

    Web of Science

    researchmap

  • Involvement of interferon regulatory factor 1 and S100C/A11 in growth inhibition by transforming growth factor beta 1 in human hepatocellular carcinoma cells

    M Miyazaki, M Sakaguchi, Akiyama, I, Y Sakaguchi, S Nagamori, NH Huh

    CANCER RESEARCH   64 ( 12 )   4155 - 4161   2004.6

     More details

    Language:English   Publisher:AMER ASSOC CANCER RESEARCH  

    Growth inhibition by transforming growth factor (TGF)-beta1 has been attributed to the induction of cyclin-dependent kinase inhibitors, among which p21/Waf1 plays a major role in many biological contexts. In the present study, two new intracellular mediators for the induction of p21/Waf1 by TGF-beta1 were identified in a human hepatocellular carcinoma cell line (JHH-5) expressing mutant-type p53. After addition of TGF-beta1 to JHH-5 cells, a marked increase of the p21/Waf1 expression preceded the inhibition of DNA synthesis. Expression of IFN regulatory factor (IRF)-1, a known transacting factor for p21/Waf1 promoter, was elevated just before or in parallel with the increase of p21/Waf1. Transduction of antisense IRF-1 inhibited the increase in p21/Waf1 in JHH-5 cells treated with TGF-beta1 and partially released the cells from the growth arrest by TGF-beta1. Expression of S100C/A11, a member of the Ca2+-binding S100 protein family, also markedly increased after addition of TGF-beta1. S100C/A11 protein was translocated to and accumulated in nuclei of TGF-beta1-treated JHH-5 cells, where p21/Waf1 was concomitantly accumulated. When a recombinant S100C/A11 protein was introduced into nuclei of JHH-5 cells, DNA synthesis was markedly inhibited in a dose-dependent manner in the absence of TGF-beta1. Prior transfection of p21/Waf1-targeted small interfering RNA efficiently blocked decrease of DNA synthesis in JHH-5 cells caused by TAT-S100C/A11 or TGF-beta1 and markedly inhibited expression of p21/Waf1 protein in the cells. These results indicate that IRF-1 and S100C/A11 mediate growth inhibition by TGF-beta1 via induction of p21/Waf1.

    DOI: 10.1158/0008-5472.CAN-03-2750

    Web of Science

    researchmap

  • Establishment of an immortalized human-liver endothelial cell line with SV40T and hTERT

    T Matsumura, M Takesue, KA Westerman, T Okitsu, M Sakaguchi, T Fukazawa, T Totsugawa, H Noguchi, S Yamamoto, DB Stolz, N Tanaka, P Leboulch, N Kobayashi

    TRANSPLANTATION   77 ( 9 )   1357 - 1365   2004.5

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Background and Aims. Liver endothelial cells (LECs) perform an essential role in important pathophysiologic functions in the liver. Establishment of a human LEC line facilitates advances in LEC research. Here, we present immortalization of human LECs using retroviral gene transfer of simian virus 40 large T antigen (SV40T) and human telomerase reverse transcriptase (hTERT). We also demonstrate excision of SV40T and hTERT with TAT-mediated Cre/loxP recombination and subsequent cell sorting.
    Methods. First, human LECs were transduced with a retroviral vector somatostatin receptor (SSR)#69 expressing SV40T and hygromycin-resistance genes flanked by a pair of loxA recombination targets. Then, cells were retrovirally superinfected with SSR#197 encoding hTERT and green fluorescent protein (GFP) cDNAs that were intervened by two loxBs. One SV40T- and hTERT-immortalized LEC clone, TMNK-1, was established and analyzed for its biologic characteristics.
    Results. The cells were hygromycin-resistant and uniformly positive for GFP expression. TMNK-1 expressed EC markers, including factor VIII, vascular endothelial growth factor receptors (flt-1, KDR/Flk-1), and CD34, showed uptake of Di-I-acetylated-low-density lipoprotein and angiogenic potential in Matrigel assays. After lipopolysaccharide treatment, TMNK-1 produced tumor necrosis factor (TNF)-alpha and interleukin (IL)-6 and exhibited increased expression of intracellular adhesive molecule-1, vascular cellular adhesive molecule-1, and VE-cadherin. After treatment with TAT-Cre recombinase fusion protein, approximately 60% of TMNK-1 was negative for GFP expression, and subsequent cell sorting of this population for GFP allowed for collection of the reverted form of TMNK-1.
    Conclusions. This study demonstrates the utility and efficiency of the reversible immortalization procedure to expand primary human LECs for basic studies.

    DOI: 10.1097/01.TP.0000124286.82961.7E

    Web of Science

    researchmap

  • Differential expression of S100C in thyroid lesions

    C Torres-Cabala, A Panizo-Santos, HC Krutzsch, H Barazi, M Namba, M Sakaguchi, DD Roberts, MJ Merino

    INTERNATIONAL JOURNAL OF SURGICAL PATHOLOGY   12 ( 2 )   107 - 115   2004.4

     More details

    Language:English   Publisher:WESTMINSTER PUBL INC  

    Identification of new potential markers that may help in the diagnosis of benign and malignant thyroid lesions is needed. By comparative 2-dimensional gel electrophoresis of microdissected cells from tumors and normal thyroid tissue, we identified a new protein, S100C, which is highly expressed in papillary carcinomas. In order to validate this finding, we investigated the immunohistochemical expression and the potential role in diagnosis of these markets in 94 specimens representing the spectrum of malignant and benign thyroid lesions. Normal thyroid tissue was evaluated in 57 specimens. Galectin-3, a marker reported as specific for malignant lesions, was also evaluated in the same lesions. S100C protein was expressed in the nuclei of normal tissue, hyperplastic nodules, and follicular adenomas and carcinomas. Papillary carcinomas showed a strong, but cytoplasmic, pattern of staining. Galectin-3 immunostaining was strongly positive in papillary carcinomas, and negative in benign lesions, confirming its value in differential diagnosis. These findings Suggest that immunohistochemical Staining of S100C could be helpful in the pathological study of thyroid lesions, especially in cases in which follicular variants of papillary carcinoma and follicular carcinoma are considered in the differential diagnosis.

    DOI: 10.1177/106689690401200203

    Web of Science

    researchmap

  • Decreased expression of REIC/Dkk-3 in human renal clear cell carcinoma

    K Kurose, M Sakaguchi, Y Nasu, S Ebara, H Kaku, R Kariyama, Y Arao, M Miyazaki, T Tsushima, M Namba, H Kumon, NH Huh

    JOURNAL OF UROLOGY   171 ( 3 )   1314 - 1318   2004.3

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Purpose: We examined the expression of REIC/Dkk-3, a possible candidate for a tumor suppressor gene, in human renal clear cell carcinoma (RCCC) cell lines and sporadic RCCC surgical specimens.
    Materials and Methods: Human RCCC cell lines (Caki-1, Caki-2, ACHN and KPK-1) and several control cell lines were used to examine the expression of REIC/Dkk-3 mRNA and characterize a newly raised antibody specific for REIC/Dkk-3 protein. Pairs of cancerous and adjacent noncancerous tissues were obtained from 20 patients with RCCC. Of them 17 and 7 cases were analyzed by real-time quantitative reverse transcriptase-polymerase chain reaction, and by Western blot analysis and/or immunohistochemical analysis, respectively.
    Results: The decreased expression of REIC/Dkk-3 mRNA and protein in human RCCC cell lines, and the specificity of the new antibody were confirmed. In a real-time quantitative reverse transcriptase-polymerase chain reaction study using 17 pairs of RCCC and adjacent normal tissues REIC/Dkk-3 mRNA levels were significantly decreased in carcinoma tissues (by 25% to approximately 95% in 15 pairs). Western blot analysis and immunohistochemistry revealed a significant decrease in REIC/Dkk-3 protein levels in 6 of the 7 and 13 of the 14 RCCC cases analyzed, respectively.
    Conclusions: The decrease in REIC/Dkk-3 mRNA and protein levels was observed irrespective of tumor grade and stage, indicating the involvement of REIC/Dkk-3 in an initial step of malignant conversion. Consequently REIC/Dkk-3 could be a new molecular target for therapeutic measures against RCCC.

    DOI: 10.1097/01.ju.0000101047.64379.d4

    Web of Science

    researchmap

  • Establishment of a highly differentiated immortalized human cholangiocyte cell line with SV40T and hTERT

    M Maruyama, N Kobayashi, KA Westerman, M Sakaguchi, JE Allain, T Totsugawa, T Okitsu, T Fukazawa, A Weber, DB Stolz, P Leboulch, N Tanaka

    TRANSPLANTATION   77 ( 3 )   446 - 451   2004.2

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Background. Cholangiocytes perform an essential role in important pathophysiologic functions in the liver. Establishment of a human cholangiocyte line facilitates advances in cholangiocyte research and clinical applications for cell therapies. Here, we describe the immortalization of human cholangiocytes using serial transfection of simian virus 40 large T (SV40T) followed by human telomerase reverse transcriptase (hTERT).
    Methods. SV40T-transduced human liver OUMS-21 cells were superinfected with a retroviral vector SSR#197 encoding hTERT and green fluorescent protein (GFP) cDNAs. Resulting cell lines were evaluated for gene expression, functional cholangiogenic characteristics in vitro and in vivo, and response to lipopolysaccharide (LPS).
    Results. One of the SV40T- and hTERT-immortalized cholangiocyte clones, MMNK-1, was established. MMNK-1 expressed cholangiocyte markers, including cytokeratin (CK)-7 and -19 and exhibited cholangiogenic tubule formation in a Matrigel assay. When transplanted into the immunodeficient mice, MMNK-1 cells developed bile duct-like structures in the spleen. After LPS treatment, MMNK-1 cells produced interleukin-6 and failed to form well-developed tubular structures in Matrigel.
    Conclusion. We have established an immortalized cholangiocyte cell line, MMNK-1, using SV40T and hTERT transduction.

    DOI: 10.1097/01.TP.0000110292.73873.25

    Web of Science

    researchmap

  • Transplantation of reversibly immortalized insulin-secreting human hepatocytes controls diabetes in pancreatectomized pigs

    T Okitsu, N Kobayashi, HS Jun, S Shin, SJ Kim, J Han, H Kwon, M Sakaguchi, T Totsugawa, M Kohara, KA Westerman, N Tanaka, P Leboulch, JW Yoon

    DIABETES   53 ( 1 )   105 - 112   2004.1

     More details

    Language:English   Publisher:AMER DIABETES ASSOC  

    Type 1 diabetes results from the destruction of insulin-producing pancreatic beta-cells by a beta-cell-specific autoimmune process. Although converting other cell types into insulin-producing cells may compensate for the loss of the beta-cell mass while evading beta-cell-specific T-cell responses, proof-of-principle of this approach in large animal models is lacking. This investigation was initiated to determine whether an insulin-producing human hepatocyte line can control diabetes when transplanted into totally pancreatectomized diabetic pigs. We established a reversibly immortalized human hepatocyte line, YOCK-13, by transferring a human telomerase reverse transcriptase cDNA and a drug-inducible Cre recombinase cassette, followed by cDNA for a modified insulin under the control of the L-type pyruvate kinase (L-PK) promoter. YOCK-13 cells produced small amounts of modified insulin and no detectable endogenous L-PK at low glucose concentrations, whereas they produced large amounts of both modified insulin and L-PK in response to high glucose concentrations. Xenotransplantation of YOCK-13 cells via the portal vein into immunosuppressed, totally pancreatectomized pigs decreased hyperglycemia and prolonged survival without adverse effects such as portal thrombosis, liver necrosis, pulmonary embolism, and tumor development. We suggest that this reversibly immortalized, insulin-secreting human hepatocyte line may overcome the shortage of donor pancreata for islet transplantation into patients with type 1 diabetes.

    DOI: 10.2337/diabetes.53.1.105

    Web of Science

    researchmap

  • Propagation of adult rat bone marrow-derived hepatocyte-like cells by serial passages in vitro

    M Miyazaki, T Masaka, Akiyama, I, E Nakashima, M Sakaguchi, NH Huh

    CELL TRANSPLANTATION   13 ( 4 )   385 - 391   2004

     More details

    Language:English   Publisher:COGNIZANT COMMUNICATION CORP  

    Previously, we found that hepatocyte growth factor receptor (c-Met)-and alpha-fetoprotein (AFP)-expressing cells were present in adult rat bone marrow, and that these cells also expressed hematopoietic stem cell markers, such as CD34, Thy-1, and c-Kit. When bone marrow cells were cultured in a hepatocyte growth medium (HGM) with HGF and EGF, colonies composed of polygonal cells resembling mature hepatocytes appeared by 2 weeks and grew very slowly because of overgrowth of stromal cells. At days 34-41, 2-mm(2) sheets of hepatocyte-like cells were cut out of their colonies by scratching with an injection needle under observation with a phase contrast microscope, transferred into wells of 24-well plates, and cultured in the HGM medium in the presence or absence of HGF and EGF. When cells reached confluence, cells were detached with trypsin and EDTA and transferred step by step into bigger culture vessels. Thus, hepatocyte-like cells were expanded 1000-fold during less than 4 months. These cells were immunocytochemically stained for albumin and also for AFP and the hematopoietic stem cell markers described above, showing characteristics of oval cells. By RT-PCR, we detected mRNAs of tryptophan-2,3-dioxygenase and tyrosine aminotransferase, markers of hepatocytes at a terminal differentiation stage. The present culture system may be useful for supply of hepatocyte resources for cell transplantation therapy.

    Web of Science

    researchmap

  • S100C/A11 is a key mediator of Ca2+-induced growth inhibition of human epidermal keratinocytes

    M Sakaguchi, M Miyazaki, M Takaishi, Y Sakaguchi, E Makino, N Kataoka, H Yamada, M Namba, NH Huh

    JOURNAL OF CELL BIOLOGY   163 ( 4 )   825 - 835   2003.11

     More details

    Language:English   Publisher:ROCKEFELLER UNIV PRESS  

    An increase in extracellular Ca2+ induces growth arrest and differentiation of human keratinocytes in culture. We examined possible involvement of S100C/A11 in this growth regulation. On exposure of the cells to high Ca2+, S100C/A11 was specifically phosphorylated at (10)Thr and (94)Ser. Phosphorylation facilitated the binding of S100C/A11 to nucleolin, resulting in nuclear translocation of S100C/A11. In nuclei, S100C/A11 liberated Sp1/3 from nucleolin. The resulting free Sp1/3 transcriptionally activated p21(CIP1/WAF1), a representative negative regulator of cell growth. Introduction of anti-S100C/A11 antibody into the cells largely abolished the growth inhibition induced by Ca2+ and the induction of p21(CIP1/WAF1). In the human epidermis, S100C/A11 was detected in nuclei of differentiating cells in the suprabasal layers, but not in nuclei of proliferating cells in the basal layer. These results indicate that S100C/A11 is a key mediator of the Ca2+-induced growth inhibition of human keratinocytes in culture, and that it may be possibly involved in the growth regulation in vivo as well.

    DOI: 10.1083/jcb.200304017

    Web of Science

    researchmap

  • 新規タンパク質リン酸化酵素BRPKの解析

    阪口 政清, 洪 梅, 片岡 健, 宮崎 正博, 許 南浩

    日本癌学会総会記事   61回   81 - 81   2002.10

     More details

    Language:Japanese   Publisher:日本癌学会  

    researchmap

  • Dynamic alteration of human beta-defensin 2 localization from cytoplasm to intercellular space in psoriatic skin

    WK Huh, T Oono, Y Shirafuji, H Akiyama, J Arata, M Sakaguchi, NH Huh, K Iwatsuki

    JOURNAL OF MOLECULAR MEDICINE-JMM   80 ( 10 )   678 - 684   2002.10

     More details

    Language:English   Publisher:SPRINGER-VERLAG  

    Defensins are cationic antimicrobial peptides with a broad spectrum. Recently human P-defensin 2 (hBD-2) has been isolated from psoriatic skin; however, its exact localization and fate have not been fully under,stood. We studied the distribution pattern of hBD-2 in skin tissues of psoriasis and other inflammatory skin diseases. In the upper spinous and granular layer of psoriasis vulgaris hBD-2 was present in the cytoplasm. In the horny layer the positive signals were in a basket-weave pattern, indicating possible accumulation of hBD-2 in the intercellular space. The similar pattern of hBD-2 distribution was observed in the lesions of nummular eczema and atopic dermatitis. hBD-2 was not detected in the section of normal elbow and knee skin. When isolated psoriatic scales were stained. hBD-2 was detected in a wrapping paper-like distribution pattern surrounding the corneocytes. In horny layer of psoriatic skin hBD-2 was closely associated or c localized with elafin, which is known to be in extracellular space, as demonstrated by double staining. Western blot analysis using cultured human keratinocytes detected hBD-2 with an expected size in the conditioned medium and in the cell lysates when stimulated with 5% FCS or IL-alpha. These results indicate that hBD-2 was synthesized and remained in cytoplasm in the upper spinous and granular layer, and then secreted into intercellular space in the horny layer. This dynamic change in hBD-2 distribution in epidermis is certainly relevant to function as an innate host defense mechanism against invading microorganisms.

    DOI: 10.1007/s00109-002-0373-z

    Web of Science

    researchmap

  • Improvement in the differentiated hepatic phenotype of immortalized human hepatocytes by adenovirus mediated p21 gene transfer

    N Kobayashi, M Sakaguchi, T Okitsu, T Totsugawa, M Maruyama, T Matsumura, T Watanabe, H Noguchi, Y Kosaka, T Fujiwara, N Tanaka

    ASAIO JOURNAL   48 ( 4 )   355 - 359   2002.7

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    The p21 molecule, a potent cyclin dependent kinase inhibitor, regulates the transition from the G1 phase to the S phase of the cell cycle and is involved in terminal cellular differentiation. The overexpression of p21 has been shown to induce differentiation in various cell lines. We have made an effort to establish a reliable human hepatocyte cell line as a source of hepatic function in bioartificial liver (BAL) therapy. In this work, we investigated the effect of p21 on the differential phenotype of simian virus 40 large T antigen (SV40Tag) immortalized human hepatocytic NKNT-3 cells. A recombinant adenoviral vector expressing a p21 gene under control of the cytomegalovirus (CMV) promoter (Ad-p21) was used to efficiently transfer genes into NKNT-3 cells. The morphologic alterations, the cell cycle progression, and the expression of p-450 associated enzymes (CYPs) were carefully examined in NKNT-3 cells that had been infected with Ad-p21. Adenovirus mediated gene delivery of p21 was efficiently achieved in NKNT-3 cells without affecting cellular structure. After Ad-p21 infection, NKNT-3 cells were GO/G1 arrested in cell cycle analysis. NKNT-3 cells that had been infected with Ad-p21 showed differentiated hepatic phenotypes in morphology and improvement in protein expression of CYP 3A4 and CYP 2C9. In the present work, we demonstrate that the exogenous expression of p21 enhances the differential phenotype of immortalized hepatocytic NKNT-3 cells.

    DOI: 10.1097/00002480-200207000-00005

    Web of Science

    researchmap

  • Reversible immortalization of human hepatocytes using the Cre/Loxp system for cell therapies

    T Totsugawa, N Kobayashi, T Okitsu, H Noguchi, T Watanabe, T Matsumura, M Maruyama, M Hikida, A Ohmori, M Sakaguchi, KA Westerman, P Leboulch, N Tanaka

    GASTROENTEROLOGY   123 ( 1 )   54 - 54   2002.7

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:W B SAUNDERS CO  

    Web of Science

    researchmap

  • Controlled expansion of human endothelial cell Populations by cre-loxP-based reversible immortalization

    H Noguchi, N Kobayashi, KA Westerman, M Sakaguchi, T Okitsu, T Totsugawa, T Watanabe, T Matsumura, T Fujiwara, T Ueda, M Miyazaki, N Tanaka, P Leboulch

    HUMAN GENE THERAPY   13 ( 2 )   321 - 334   2002.1

     More details

    Language:English   Publisher:MARY ANN LIEBERT INC PUBL  

    Endothelial cells (ECs) play multiple physiological functions and are central to many pathological processes. Various biological studies as well as cell and gene therapy applications would benefit substantially from a procedure that would result in the expansion in culture of large numbers of highly differentiated human ECs. Here, we report the amplification in vitro of human EC populations, which occurred during the first phase of reversible immortalization resulting from the retroviral transfer of an oncogene that was subsequently excised by Cre-loxP-mediated site-specific recombination. Human umbilical vein endothelial cells (HUVECs) and human liver sinusoidal endothelial cells (HLSECs) were transduced with a retroviral vector that expresses the simian virus 40 large T (SV40T) gene flanked by positive and negative selectable markers and a pair of loxP recombination targets. Transduced HUVECs and HLSECs yielded clones with greatly extended life spans, referred to as HNNT-1 and HNNT-2 cells, respectively. HNNT-1 and HNNT-2 cells showed morphological characteristics of ECs and were maintained in culture up to population doubling level (PDL) 80 for HNNT-1 and PDL 65 for HNNT-2 cells. HNNT-1 and HNNT-2 cells were not tumorigenic when transplanted into severe combined immunodeficiency mice and were sensitive to ganciclovir as well as G418. Both cell clones expressed EC markers, which include factor VIII, VEGF receptors (Flt-1 and KDR/Flk-1), and CD34, and endocytosed acetylated low-density lipoproteins. Formation of capillary-like structures in a Matrigel assay was observed with HNNT-1 and HNNT-2 cells until at least PDL 50. Complete elimination of the transferred SV40T gene was achieved in virtually 100% of HNNT-1 and HNNT-2 cells after infection with a recombinant adenovirus expressing the Cre recombinase fused to a nuclear localization signal and subsequent selection with G418. Reverted cells maintained their differentiated EC phenotype. This study extends the utility of the reversible immortalization procedure and provides a means to expand primary human ECs of various sources for basic studies and possible cell and gene therapies.

    DOI: 10.1089/10430340252769833

    Web of Science

    researchmap

  • Lentiviral transfer of the LacZ gene into human endothelial cells and human bone marrow mesenchymal stem cells

    T Totsugawa, N Kobayashi, T Okitsu, H Noguchi, T Watanabe, T Matsumura, M Maruyama, T Fujiwara, M Sakaguchi, N Tanaka

    CELL TRANSPLANTATION   11 ( 5 )   481 - 488   2002

     More details

    Language:English   Publisher:COGNIZANT COMMUNICATION CORP  

    Because one of the attractive characteristics of human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors is that it can infect even nondividing cells, a lentivirus-mediated gene delivery system is currently being paid a great deal of attention as an innovative tool for gene transfer into target cells. The purpose of the work was to investigate the efficacy of lentiviral transfer of the LacZ gene into human umbilical vein endothelial cells (HUVECs) and human bone marrow mesenchymal stem cells (HMSCs) in vitro. For the present study, a vesicular stomatitis virus G-protein (VSV-G)-pseudotyped lentiviral vector encoding the E. coli LacZ gene tagged with nuclear localization signal (NLS) was generated in 293T cells by means of the three-plasmid system. The resulting lentiviral vector, LtV-NLS/LacZ, was allowed to infect HUVECs and HMSCs. Approximately 70% of HUVECs were positive for LacZ expression and 50% of HMSCs showed LacZ activity. There was no significant difference in transduction efficacy between early and late-passage phases in both cells. LtV-NLS/LacZ-transduced HUVECs showed gene expression of endothelial markers including CD34 and flt-1 and KDR/flk-1 of vascular endothelial growth factor (VEGF) receptors and had angiogenic potential as efficiently as primarily cultured HUVECs in a Matrigel assay. These findings provide evidence that lentiviral vectors are efficient tools for gene transfer and expression in human endothelial cells and stem cells that could be useful for tissue engineering.

    Web of Science

    researchmap

  • Transduction of immortalized human hepatocytes with p21 to enhance differentiated phenotypes

    T Kunieda, N Kobayashi, M Sakaguchi, T Okitsu, T Totsugawa, T Watanabe, T Matsumura, M Maruyama, H Noguchi, M Takesue, N Shibata, K Ohmoto, T Fujiwara, S Yamamoto, N Tanaka

    CELL TRANSPLANTATION   11 ( 5 )   421 - 428   2002

     More details

    Language:English   Publisher:COGNIZANT COMMUNICATION CORP  

    We previously constructed an immortal human hepatocyte line NKNT-3 with a simian virus 40 T antigen (SV40T) to develop cell-based biological therapies. p21 is a molecule that regulates the transition from the G, phase to the S phase of the cell cycle. Investigators have demonstrated that overexpression of p21 induces differentiation in various cell lines. In the current study we examined the effect of p21 on differentiated phenotypes of SV40T-immortalized NKNT-3 cells. A replication-deficient adenovirus vector expressing a human wild-type p21 cDNA under the control of the CMV promoter (Ad5CMVp21) and a human wild-type p21 protein fused to the protein transduction domain from the human immunodeficiency virus (HIV) TAT protein (TAT/p21) were utilized to achieve efficient delivery of p21 into NKNT-3 cells. Morphological alterations, cell cycle progression, and expression of albumin and p-450 associated enzymes (CYPs) 3A4 and 2C9 were evaluated in NKNT-3 cells treated with Ad5CMVp21 and TAT/p21. Efficient adenovirus-based p21 transfer and TAT-mediated p21 protein delivery were confirmed in NKNT-3 cells in an immunofluorescence study and Western blotting analysis. Transduction of NKNT-3 cells with p21 predominantly arrested the cell cycle at the G, checkpoint, resulting in differentiated hepatic phenotypes in morphology and improvement in protein expression of albumin, CYP 3A4, and CYP C29. We here show that exogenous expression of p21 augments cellular differentiation in immortalized human NKNT-3 cells.

    Web of Science

    researchmap

  • レンチウイルスベクターシステム樹立に伴う安全配慮.

    都津川敏範, 小林直哉, 丸山昌伸, 小坂芳和, 興津 輝, 荒田 尚, 阪口政清, 藤原俊義, 倉林 譲, 田中紀章

    Organ Biology   9,4,357-364   2002

     More details

  • Reactivation of liver-specific gene expression in an immortalized human hepatocyte cell line by introduction of the human HNF4 alpha 2 gene

    Y Inoue, M Miyazaki, T Tsuji, M Sakaguchi, K Fukaya, NH Huh, M Namba

    INTERNATIONAL JOURNAL OF MOLECULAR MEDICINE   8 ( 5 )   481 - 487   2001.11

     More details

    Language:English   Publisher:PROFESSOR D A SPANDIDOS  

    An immortalized human hepatocyte cell line (OUMS-29) was established from fetal liver by transfection with the SV-40 large T antigen gene that has certain liver-specific functions such as albumin production and enzyme activities of CYP1A1, 1A2, and 2E1. To make OUMS-29 cells express other liver-specific functions, the human hepatocyte nuclear factor 4 alpha2 (HNF4 alpha2) gene was introduced into the cells, because this gene was found to be markedly down-regulated. The transduced HNF4 alpha2 was overexpressed in the nuclei of the transfected cells, and its DNA-binding activity was also detected. The liver-specific genes such as apolipoprotein AI, CII, CIII, blood coagulation factor X, alpha1-antitrypsin, and HNF1 alpha were up-regulated. Thus, this cell line is expected to be a useful tool for studying the differentiated human hepatocyte, functions.

    Web of Science

    researchmap

  • Adenovirus-mediated p21 gene transfer enhances differentiated hepatic phenotypes of immortalized human hepatocytes.

    N Kobayashi, H Noguchi, T Totsugawa, T Watanabe, T Matsumura, M Maruyama, T Matsumoto, T Fujimara, N Tanaka, M Sakaguchi

    HEPATOLOGY   34 ( 4 )   305A - 305A   2001.10

     More details

    Language:English   Publishing type:Research paper, summary (international conference)   Publisher:W B SAUNDERS CO  

    Web of Science

    researchmap

  • Construction of a differentiated human hepatocyte cell line expressing the herpes simplex virus-thymidine kinase gene

    N Kobayashi, M Miyazaki, KA Westerman, H Noguchi, M Sakaguchi, T Totsugawa, T Watanabe, T Matsumura, T Fujiwara, P Leboulch, N Tanaka, M Namba

    ASAIO JOURNAL   47 ( 5 )   476 - 480   2001.9

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    Transient support using a hybrid artificial liver (HAL) device is a promising treatment for the patients with acute liver failure. Primary human hepatocytes are an ideal source for HAL therapy; however, the number of human livers available for hepatocyte isolation is limited by competition for use in whole organ transplantation. To overcome this problem, we previously established a highly differentiated human fetal hepatocyte cell line OUMS-29. Considering the potential risk when using these genetically engineered cells in humans, additional safeguards should be added to make the cells more clinically useful. In this work, the herpes simplex virus thymidine kinase (HSVtk) gene was retrovirally introduced into OUMS-29 cells. One of the HSVtk-expressed clones, OUMS-29/thymidine kinase (TK), grew in chemically defined serum free medium and expressed the genes of albumin, asialoglycoprotein receptor, glutamine synthetase, glutathione-S-transferase pi, and blood coagulation factor X. In vitro sensitivity of the cells to ganciclovir was evaluated. Intrasplenic transplantation of 50 X 10(6) OUMS-29/TK cells prolonged the survival of 90% hepatectomized rats compared with medium injection alone (control). In the present study, we have established highly differentiated immortalized human hepatocytes with tight regulation. The cells may be clinically useful for HAL treatment.

    DOI: 10.1097/00002480-200109000-00016

    Web of Science

    researchmap

  • Expansion of human hepatocyte populations by a retroviral gene transfer of simian virus 40 large T antigen

    N Kobayashi, KA Westerman, T Taguchi, M Sakaguchi, T Fujiwara, H Urata, N Kishimoto, N Hayashi, S Nakaji, T Murakami, P Leboulch, N Tanaka

    ASAIO JOURNAL   47 ( 5 )   481 - 485   2001.9

     More details

    Language:English   Publisher:LIPPINCOTT WILLIAMS & WILKINS  

    A hybrid artificial liver (HAL) could be used to treat acute liver failure or to serve as a temporary support until orthotopic liver transplantation is available. Primary human hepatocytes are ideal as a source of hepatic function in a HAL device. However, the worldwide shortage of human livers available for hepatocyte isolation severely limits this form of therapy. A possible alternative is to use a tightly regulated cell line that can be economically grown in culture to have differentiated liver function. In this work, human hepatocytes were immortalized with a retroviral vector SSR#69 expressing the genes of simian virus 40 large T antigen and herpes simplex virus-thymidine kinase. One of the resulting clones, NKNT-3, showed the gene expression of differentiated liver function and were sensitive to the antiviral agent ganciclovir. When transplanted into the spleen of rats subjected to 90% hepatectomy, NKNT-3 cells prolonged the survival of 90% hepatectomized rats. The cells provide the advantages of unlimited availability, sterility, uniformity, and freedom from pathogens. This work represents a potential novel strategy for resolving the organ shortage that currently limits the use of primary human hepatocytes to develop a HAL.

    DOI: 10.1097/00002480-200109000-00017

    Web of Science

    researchmap

  • レンチウイルスベクターによるヒト血管内皮細胞への効果的な遺伝子導入.

    都津川敏範, 小林直哉, 丸山昌伸, 興津 輝, 野口洋文, 松村年久, 渡辺剛正, 阪口政清, 藤原俊義, 田中紀章

    Organ Biology   2001

     More details

  • 【臓器移植と医工学 その現況と展望】 急性肝不全に対する不死化ヒト肝細胞の移植

    小林 直哉, 野口 洋文, 田中 紀章, 深谷 憲一, 阪口 政清, 井上 祐介, 宮崎 正博, 難波 正義

    今日の移植   13 ( 3 )   216 - 221   2000.5

     More details

    Language:Japanese   Publisher:(株)日本医学館  

    researchmap

▼display all

Presentations

  • Xylitol mediates selective cancer death through regulation of the glutathione level

    Nahoko Tomonobu, Yuma Gohara, Rie Kinoshita, Masakiyo Sakaguchi

    2021.10.1 

     More details

    Event date: 2021.9.30 - 2021.10.2

    Language:Japanese   Presentation type:Poster presentation  

    researchmap

  • Newly developed anti-S100A8 and A9 heterodimer monoclonal antibody for treatment of cancer metastasis and idiopathic pulmonary fibrosis

    2021.9.3 

     More details

    Event date: 2021.9.2 - 2021.9.3

    Language:Japanese   Presentation type:Oral presentation (general)  

    researchmap

  • Alcohol Extracts from Plastic Wrap induces Cytotoxic Effect on Culture cells

    2021.9.3 

     More details

    Event date: 2021.9.2 - 2021.9.3

    Language:Japanese   Presentation type:Poster presentation  

    researchmap

  • Phosphorylated SARM1 is involved in the pathological process of Parkinson’s disease

    2021.9.3 

     More details

    Event date: 2021.9.2 - 2021.9.3

    Language:Japanese   Presentation type:Oral presentation (general)  

    researchmap

  • Xylitol is a potent anti-cancer monosaccharide that induces cancer-selective cell death by regulating the intracellular glutathione Invited

    2021.9.2 

     More details

    Event date: 2021.9.2 - 2021.9.3

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    researchmap

  • がんの転移制御を目指して Invited

    阪口政清

    岡山肺癌基礎研究会  2021.6.11 

     More details

    Event date: 2021.6.11

    Language:Japanese   Presentation type:Oral presentation (invited, special)  

    Venue:岡山市  

    researchmap

  • HNF4αとCREBHによる肝臓のFsp27bの相乗的な発現制御

    笠野 一郎, 瀧澤 将行, 岩崎 若菜, 佐々木 翔太, 濱田 夢芽, 守本 葵, 阪口 政清, GONZALEZ Frank J, 井上 裕介

    第44回日本分子生物学会年会  2020.12.2 

     More details

    Event date: 2020.12.2 - 2020.12.4

    Language:Japanese   Presentation type:Poster presentation  

    Venue:オンライン  

    researchmap

  • HNF4γ2に相互作用するタンパク質の探索

    江原 伸哉, 佐々木 翔太, 浦部 瑞穂, 前田 つかさ, 鈴木 淳子, 入江 亮太, 鈴木 正則, 外丸 靖浩, 阪口 政清, FRANK Gonzalez J, 井上 裕介

    第44回日本分子生物学会年会  2020.12.2 

     More details

    Event date: 2020.12.2 - 2020.12.4

    Language:Japanese   Presentation type:Poster presentation  

    Venue:オンライン  

    researchmap

  • ロテノン誘発パーキンソニズムの病態形成におけるSARM1リン酸化制御の意義

    村田 等, 越智 俊樹, 山本 健一, 木下 理恵, 阪口 政清

    第43回日本分子生物学会年会  2020.12.2 

     More details

    Event date: 2020.12.2 - 2020.12.4

    Language:Japanese   Presentation type:Poster presentation  

    Venue:オンライン  

    researchmap

  • The secretory S100A11 activates fibroblasts surrounding tumor that enhances cell motility in pancreatic cancer.

    Gohara Yum, Mitsui Yosuke, Tomonobu Nahoko, Kinoshita Rie, Yamamoto Kenich, Yamauchi Akira, Yamamura Masahiro, Kondo Eisaku, Toyooka Shinich, Nasu Yasutomo, Murata Hitoshi, Masakiyo Sakaguchi

    2020.12.2 

     More details

    Event date: 2020.12.2 - 2020.12.4

    Language:Japanese   Presentation type:Poster presentation  

    researchmap

  • Eextracellular S100A11 upregulates mobility of pancreatic cancer cells through activation of surrounding fibroblasts

    2020.10.1 

     More details

    Event date: 2020.10.1 - 2020.10.3

    Language:Japanese   Presentation type:Poster presentation  

    researchmap

  • HSP90 inhibition suppresses not only cell proliferation but also chemotaxis in esophageal squamous cell carcinoma

    2020.10.1 

     More details

    Event date: 2020.10.1 - 2020.10.3

    Language:Japanese   Presentation type:Oral presentation (general)  

    researchmap

  • NOX4 on lymphatic endothelial cells (LEC;plays an important;role on;the migration of;pancreatic cancer cells

    2020.10.1 

     More details

    Event date: 2020.10.1 - 2020.10.3

    Language:Japanese   Presentation type:Oral presentation (general)  

    researchmap

  • Study of the xylitol-mediated selective cancer death and evaluation of the anticancer effect of xylitol in vivo

    Nahoko Tomonobu, Yuma Gohara, Rie Kinoshita, Masakiyo Sakaguchi

    2020.10.1 

     More details

    Event date: 2020.10.1 - 2020.10.3

    Language:Japanese   Presentation type:Poster presentation  

    researchmap

  • Heat shock protein 90 inhibition suppresses not only proliferation but also migration towards lysophosphatidic acid and Sphingosine-1-phosphate in esophageal squamous cell carcinoma International conference

    Masahiro Yamamura, Akira Yamauchi, Naoki Katase, Masakiyo Sakaguchi, Yosuke Katata, Fuminori Sano, Hiroaki Tanioka, Makoto Okawaki, Takeshi Nagasaka, Yoshiyuki Yamaguchi

    AACR Annual Meeting 2020  2020.4.24 

     More details

    Event date: 2020.4.24 - 2020.4.29

    Language:English  

    researchmap

  • S100 family protein: its role in inflammation associated with aging, Specific title: Prevention of cancer metastasis on the basis of identification of novel S100 protein sensor receptors Invited International conference

    National Symposium and Workshop in Anti-Aging Medicine (NASWAAM)  2020.2.7 

     More details

    Event date: 2020.2.7 - 2020.2.9

    Language:English   Presentation type:Oral presentation (general)  

    researchmap

  • Prevention of Aging Process: Biomolecular insight, Specific title: A Novel Tumor Suppressor, REIC/Dkk-3 Gene Identified by Our In Vitro Transformation Model of Normal Human Fibroblasts Works as a Potent Therapeutic Anti-tumor Agent Invited International conference

    National Symposium and Workshop in Anti-Aging Medicine (NASWAAM)  2020.2.7 

     More details

    Event date: 2020.2.7 - 2020.2.9

    Language:English   Presentation type:Oral presentation (invited, special)  

    researchmap

  • 肺腺がん細胞における核内受容体HNF4αの機能解析と標的遺伝子の同定

    亀井めぐみ, 濱田真輝, 阪口政清, 井上裕介

    第42回日本分子生物学会年会  2019.12.5 

     More details

    Event date: 2019.12.3 - 2019.12.6

    Presentation type:Poster presentation  

    Venue:福岡市  

    researchmap

  • Development of exSSSRs(extracellular S100 soil sensor receptors)-Fc fusion proteins and S100A8/A9 antibody for suppression of cancer metastasis International conference

    Rie Kinoshita, Hiroki Sato, Akira Yamauchi, Yuta Takahashi, Yusuke Inoue, I Wayan Sumardika, Youyi Chen, Nahoko Tomonobu, Kota Araki, Kazuhiko Shien, Shuta Tomida, Hidejiro Torigoe, Kei Namba, Eisuke Kurihara, Yusuke Ogoshi, Hitoshi Murata, Ken-ichi Yamamoto, Junichiro Futami, Endy Widya PutrantoI, I Made Winarsa Ruma, Hiromasa Yamamoto, Junichi Soh, Toshihiko Hibino, Masahiro Nishibori, Eisaku Kondo, Shinichi Toyooka, Masakiyo Sakaguchi

    2019.11.6 

     More details

    Event date: 2019.11.6 - 2019.11.8

    Language:English   Presentation type:Poster presentation  

    researchmap

  • Melanoma cell adhesion molecule (MCAM) induces dissemination of melanoma upon S100A8/A9 binding

    2019.9.28 

     More details

    Event date: 2019.9.26 - 2019.9.28

    Presentation type:Poster presentation  

    researchmap

  • Novel therapeutic approach based on S100A8/A9-mediated organ tropic cancer metastasis

    2019.9.27 

     More details

    Event date: 2019.9.26 - 2019.9.28

    Presentation type:Poster presentation  

    researchmap

  • JNKによるリン酸化を介したSARM1のNAD+代謝と軸索変性への寄与 Invited

    村田 等, 阪口政清

    第92回日本生化学会大会  2019.9.18 

     More details

    Event date: 2019.9.18 - 2019.9.20

    Presentation type:Symposium, workshop panel (nominated)  

    Venue:横浜市  

    researchmap

  • Requlation of SARM1-NAD+ cleavage activity and mitochondrial function through phosphorylation by JNK

    2019.7.25 

     More details

    Event date: 2019.7.25 - 2019.7.28

    Presentation type:Poster presentation  

    researchmap

  • 新規HNF4γバリアントによる肝機能の誘導

    佐々木 翔太, 浦部瑞穂, 前田つかさ, 鈴木淳子, 入江亮太, 阪口政清, Frank J. Gonzalez, 井上裕介

    第  2018.11.30 

     More details

    Event date: 2018.11.28 - 2018.11.30

    Presentation type:Poster presentation  

    researchmap

  • 分泌性S100A11-受容体RAGEシグナルを介した膵臓がん周辺微小環境における間質線維芽細胞の増殖誘導の解明

    山本 健一, 高松 仁志, 友信 奈保子, 光井 洋介, 二見 淳一郎, 木下 理恵, 村田 等, 阪口 政清

    第41回日本分子生物学会年会  2018.11.30 

     More details

    Event date: 2018.11.28 - 2018.11.30

    Language:Japanese   Presentation type:Poster presentation  

    Venue:横浜市  

    researchmap

  • JNK-mediated phosphorylation of SARM1 regulates NAD cleavage activity to inhibit mitochondrial respiration

    2018.11.30 

     More details

    Event date: 2018.11.28 - 2018.11.30

    Presentation type:Poster presentation  

    researchmap

  • Development of novel biologics to prevent lung tropic cancer metastasis. Invited International conference

    Masakiyo Sakaguchi

    The First International Symposium on Immunology and Cancer in Okayama / 13th URA International Symposium  2018.10.31 

     More details

    Event date: 2018.10.31

    Language:English   Presentation type:Oral presentation (invited, special)  

    researchmap

  • Development of a novel S100A8/A9 neutralizing monoclonal antibody for suppression of cancer metasasis.

    2018.9.28 

     More details

    Event date: 2018.9.27 - 2018.9.29

    Language:English   Presentation type:Poster presentation  

    researchmap

  • Tumor-suppressive effect of LRIG1 in non-small cell lung cancer harboring mutant EGFR

    Hidwejiro Torigoe, Hiromasa Ymamoto, Masakiyo Sakaguchi, Kei Namba, Hiroki Sato, Kazunori Shien, Ken Suzawa, Junichi Soh, Shuta Tomida, Kazunori Tsukuda, Shinichiro Miyoshi, Shinichi Toyooka

    The 77th Annual Meeting of the Japanese Cancer Association  2018.9.28 

     More details

    Event date: 2018.9.27 - 2018.9.29

    Language:English   Presentation type:Oral presentation (general)  

    researchmap

  • Biological role of PODXL1 in invasion and metastasis of Pancreatic ductal adenocarinoma.

    Eisaku Kondo, Masakiyo Sakaguchi, Hidekazu Iioka, Ken Saito

    The 77th Annual Meeting of the Japanese Cancer Association  2018.9.27 

     More details

    Event date: 2018.9.27 - 2018.9.29

    Language:English   Presentation type:Oral presentation (general)  

    researchmap

  • 分泌性S100A11-受容体RAGEシグナルに着眼した膵がん間質増大のメカニズムの解明分泌性S100A11-受容体RAGEシグナルに着眼した膵がん間質増大のメカニズムの解明

    本健一, 高松仁志, 光井洋介, 木下理恵, 村田 等, 二見淳一郎, 山本靖彦, 西堀正洋, 豊岡伸一, 阪口政清

    第22回日本がん免疫学会総会  2018.8.3 

     More details

    Event date: 2018.8.1 - 2018.8.3

    Language:Japanese   Presentation type:Oral presentation (general)  

    researchmap

  • 膵がん進展に導く膵がん細胞−間質線維芽細胞クロストークを介在する分泌性 S100A11−受容体RAGE連携の役割

    光井洋介, 山本健一, Sumardika I Wayan, 木下理恵, 村田等, 二見淳一郎, 高松仁, 山本靖彦, 西堀正洋, 豊岡伸一, 渡部昌実, 那須保友, 阪口政清

    第22回日本がん免疫学会総会  2018.8.2 

     More details

    Event date: 2018.8.1 - 2018.8.3

    Language:Japanese   Presentation type:Poster presentation  

    researchmap

  • Novel biologics to prevent cancer metastasis Invited International conference

    Masakiyo Sakaguchi

    第9回International DAMPs and Alarmins Symposium  2019 

     More details

    Language:Japanese  

    researchmap

  • s100a8/A9タンパク質と臓器指向性転移

    第28回創薬・薬理フォーラム岡山  2017 

     More details

  • Therapeutic Strategies for suppression of cancer metastasis on the basis of finding novel S100A8/A9 receptors

    2017 

     More details

  • Development of novel biologics for cancer metastasis via prevention of extracellular S100A8/A9 function

    The 76th Annual Meeting of the Japanese Cancer Association  2017 

     More details

  • Crumbs3 promotes metastasis of colon cancer by regulating focal adhesion components

    The 76th Annual Meeting of the Japanese Cancer Association  2017 

     More details

  • Identification of novel receptors for pro-inflammatory S100A8/A9 protein and its role(s) in cancer metastasis

    2017 

     More details

  • SARM1 induces neuronal cell death by inhibition of mitochondrial respiration

    2017 

     More details

  • S100タンパク質を基軸とした乾癬病態増悪の分子メカニズムとその制御

    第16回皮膚科スプリングセミナー  2017 

     More details

  • S100A8を標的としたがん転移制御法の開発

    日本組織培養学会第90回大会  2017 

     More details

  • 転移先臓器を感知する受容体

    日本組織培養学会第90回大会  2017 

     More details

  • Anti-glioma mechanism of the Ad-REIC vector with the super gene expression system.

    2017 

     More details

  • S100A8/A9 とその受容体との結合遮断を目指した転移抑制タンパク質製剤の開発

    第76回日本癌学会学術総会  2017 

     More details

  • ミトコンドリア呼吸鎖複合体への作用を介したSARM1の神経細胞死誘導

    日本組織培養学会第90回大会  2017 

     More details

  • A super gene expression system enhances the anti-glioma effects of adenovirus-mediated REIC/Dkk-3 gene therapy.

    2017 

     More details

  • 核内受容体HNF4γによる肝がん細胞の再分化誘導

    第39回日本分子生物学会年会  2017 

     More details

  • ゲノムと遺伝情報-5) RNAプロセシング、輸送、翻訳、非コードRNA

    第39回日本分子生物学会年会  2017 

     More details

  • Crumbs3 は焦点接着構成因子を制御することで大腸癌の転移を促進する

    第76回日本癌学会学術総会  2017 

     More details

  • 健常者及びパーキンソン病患者iPS細胞由来の神経細胞を用いたミトコンドリア機能解析

    第39回日本分子生物学会年会  2017 

     More details

  • HNF4αによるmiRNAを介した遺伝子発現制御機構の解析

    第39回日本分子生物学会年会  2016 

     More details

  • Development of a novel biologics for suppression of S100A8/A9-induced cancer metastasis

    The 76th Annual Meeting of the Japanese Cancer Association  2016 

     More details

  • Beta3 integrin cDNA-transduced human keratinocytes grow far better on fibrin gels as compared to normal human keratinocytes

    2016 

     More details

  • Extract of Cordyceps militaries inhibits angiogenesis and suppresses tumor growth of human malignant melanoma cells

    2016 

     More details

  • Functional Interplay between the Leukotriene B4 receptor BLT1 and RAGE

    6th international conference on Phospholipase A2 and Lipid Mediators(PLM2015)  2016 

     More details

  • Regulation of Cellular Stress Response By Mitochondrial Kinase Pink1

    2016 

     More details

  • Interaction between Pancreatic cancer cells and MSC for cancer progression

    The 76th Annual Meeting of the Japanese Cancer Association  2016 

     More details

  • The cell polarity regulator Crumbs3 promotes adenocarcinoma metastasis through the regulation of glycolipid dynamics

    The 76th Annual Meeting of the Japanese Cancer Association  2016 

     More details

  • PINK1 regulation of mitochondrial homeostasis and cell survival.

    the 2016 World Congress on In Vitro Biology  2016 

     More details

  • 肺動脈性肺高血圧症におけるRAGEシグナルの亢進

    第4回日本肺循環学会・第3回日本肺高血圧学会 合同学術集会  2016 

     More details

  • 冬虫夏草抽出溶液による悪性メラノーマ細胞の増殖と血管新生の抑制

    日本組織培養学会第89回大会  2016 

     More details

  • Beta3インテグリンcDNA導入ヒトケラチノサイトは正常ヒトケラチノサイトに比べてフィブリンゲル上で細胞増殖が増加する

    日本組織培養学会第89回大会  2016 

     More details

  • 癌転移抑制を目指した新規タンパク質製剤の開発

    第75回日本癌学会学術総会  2016 

     More details

  • 浸潤性膵管癌におけるがん-間質相互反応の増殖・浸潤における役割

    第75回日本癌学会学術総会  2016 

     More details

  • 細胞極性制御因子Crumbs3 は糖脂質動態を制御することで腺癌の転移を促進する

    第75回日本癌学会学術総会  2016 

     More details

  • 核内受容体HNF4γの機能解析

    第39回日本分子生物学会年会  2016 

     More details

  • 神経細胞死・軸索変性に関与するSARM1のリン酸化制御

    第39回日本分子生物学会年会  2016 

     More details

  • NOD2 inflammasome is up-regulated in the skin with atopic dermatitis.

    日本研究皮膚科学会 第40回年次学術大会・総会  2015 

     More details

  • Identification of a novel binding protein playing a critical role in HER2 activation in lung cancer cells

    AACR Annual Meeting 2015  2015 

     More details

  • RegulationofREIC/Dkk-3expressionbyTNF-alphaanditseffecton normal human keratinocytes

    2015 

     More details

  • NRF2 pathway activation as a target to counteract mitochondrial dysfunction in Parkinson's disease

    2015 

     More details

  • TNF-αによるREIC/Dkk-3の発現制御とケラチノサイトの挙動への影響

    日本組織培養学会第88回大会  2015 

     More details

  • TIR/BB-loop mimetic AS-1 Inhibits Proliferation of Pulmonary Artery Smooth Muscle Cells from Patients with Pulmonary Arterial Hypertension

    第79回日本循環器学会  2015 

     More details

  • S100A8/A9 induces skin inflammation and keratinocyte proliferation via neuroplasatin-beta/EMMPRIN heterodimer receptor in atopic dermatitis

    日本研究皮膚科学会 第40回年次学術大会・総会  2015 

     More details

  • S100A8 promoted Th1-related gene expression and S100A9 induced Th2 differentiation via activation of GATA-3

    日本研究皮膚科学会 第40回年次学術大会・総会  2015 

     More details

  • Mechanism of S100A9 induction in the skin with atopic dermatitis―involvement of NOD2 inflammasome

    日本研究皮膚科学会 第40回年次学術大会・総会  2015 

     More details

  • Regulation of PINK1 expression through the NRF2 transcription factor under mitochondrial stress conditions

    PINK1-Parkin Signalling in Parkinson’s Disease and Beyond  2014 

     More details

  • Novel functional mutations in the transmembrane domain of HER2 gene in familial and sporadic lung adenocarcinomas

    2014 

     More details

  • Novel HER2 mutations in transmembrane dmain result in constitutive autophosphorylation of HER2

    2014 

     More details

  • 新開発超高効率遺伝子発現プラスミドベクターによる抗体大量産出技術の確立

    大学研究力強化支援情報交換ワークショップ  2014 

     More details

  • IgG4関連疾患:病態形成機序ならび腫瘍との関連性

    第17回がんと免疫セミナー  2014 

     More details

  • 炎症を感知する新規内因性リガンドセンサ―の作動機構解明とがん進展における役割

    新学術領域平成25年度第4回領域班会議  2014 

     More details

  • Critical role of Cytokeratin-19 in an oncogenic activation of HER2

    2014 

     More details

  • S100タンパク質によるケラチノサイトの増殖、死の分子制御機構、阪口政清

    資生堂新領域研究センター特別講演  2014 

     More details

  • 家族性・孤発性肺腺癌におけるHER2 膜貫通領域の新規遺伝子変異

    第73回日本癌学会学術総会  2014 

     More details

  • 新規HER2 膜貫通部領域遺伝子変異の機能解析

    第73回日本癌学会学術総会  2014 

     More details

  • HNF4γの機能解析

    第37回日本分生生物学会年会  2014 

     More details

  • ロイコトリエンB4第一受容体BLT1とRAGEの相互作用

    第87回日本生化学会大会  2014 

     More details

  • 高機能REIC 遺伝子発現アデノウイルスベクターの開発

    岡山大学機能強化戦略プロジェクト-難治固形がんの遺伝子治療- 第2回シンポジウム  2014 

     More details

  • HER2がんマーカー,サイトケラチン19 (CK19) 機能の新展開― HER2活性化におけるCK19 の役割―

    第73回日本癌学会学術総会  2014 

     More details

  • Histidine-rich glycoprotein prevents septic lethality through neutrophil regulation

    Sepsis Symposium at Pasteur Institute in Paris  2014 

     More details

  • Identification of RAGE-coupled membran adaptor that modulates RAGE-triggered signaling pathway

    2013年アメリカ細胞生物学会総会  2013 

     More details

  • Regulation of Cellular Stress Response By Mitochondrial Kinase Pink1

    日本組織培養学会第86回大会  2013 

     More details

  • S100A8/A9 targets oncogenic survival pathway via RAGE that critically enhanced by a RAGE co-receptor. 多機能受容体RAGE の共役受容体の発見とその意義

    第72回日本癌学会学術総会  2013 

     More details

  • 膿疱性乾癬の病態解明とその対策に向けて - S100A8およびS100A9タンパク質の新規受容体の探索とその機能解析-

    稀少難治性皮膚疾患に関する調査研究班平成24年度第1回総会  2013 

     More details

  • 多機能受容体RAGEの共役受容体の発見とその意義

    新学術領域研究「自然炎症」+「脂質マシナリー」合同若手ワークショップ  2013 

     More details

  • S100タンパク質機能解析の新展開―分泌性S100タンパク質とその受容体ファミリーによる「炎症―腫瘍進展」応答ネットワーク形成の分子機構の解明とその制御

    徳島文理大学大学院特別講演  2013 

     More details

  • SARM1and TRAF6 bind to and stabilizes PINK1 on the outer membrane of depolarized mitochondria for mitophagy through interaction with SARM1

    第36回日本分子生物学会年会  2013 

     More details

  • TRAF6 and SARM1 regulate PINK1 ubiquitination and stabilization on

    第1回国際ミトコンドリア学術集会  2013 

     More details

  • 肝臓特異的HNF4α欠損マウスにおけるHNF4γの機能解析

    第36回日本分子生物学会年会  2013 

     More details

  • TRAF6 ubiquitinates and stabilizes PINK1 on the outer membrane of depolarized mitochondria through interaction with SARM1

    2013年アメリカ細胞生物学会総会  2013 

     More details

  • Analysis of Pseudolysogenic Lifecycle of Clostridium botulinum Phage c-st, Based on Comparative Genomics of Wild-type and Long-term-incubated Mutants Phages

    2012 

     More details

  • Identification of novel receptors for pro-inflammatory S100A8/A9 proteins and their potential

    2012 

     More details

  • Development of RAGE-inhibitor delivered into cells

    2012 

     More details

  • Analysis of Pseudolysogenic Lifecycle of Clostridium botulinum Phage c-st, Based on Comparative Genomics of Wild-type and Long-term-incubated Mutants Phages

    2012 

     More details

  • TRAF6 Ubiquitinates and Stabilizes PINK1 on Damaged Mitochondria Through Interaction with SARM1

    2012 

     More details

  • DOCK7 is a critical regulator of the RAGE-Cdc42 signaling axis that induces formation of dendritic pseudopodia in human cancer cells

    2012 

     More details

  • 膿疱性乾癬の病態解明とその対策に向けて - S100A8およびS100A9タンパク質の新規受容体の探索とその機能解析-

    稀少難治性皮膚疾患に関する調査研究班平成24年度第1回総会  2012 

     More details

  • A Novel Tumor Suppressor, REIC/Dkk-3 Gene Identified by our in Vitro Transformation Model of Normal Human Fibroblasts Works as a Potent Therapeutic Anti-tumor Agent

    2012 World Congress on In Vitro Biology  2012 

     More details

  • 細胞内導入型RAGE阻害剤の開発

    日本組織培養学会第85回大会  2012 

     More details

  • 細胞遊走に関わるRAGE膜直下信号伝達機構の解析―RAGE細胞質領域結合タンパク質Dock7の発見―

    第35回日本分子生物学会年会  2012 

     More details

  • Identification of novel receptors for pro-inflammatory S100A8/A9 proteins and their roles inmelanoma metastasis

    IEIIS2012. Homeostatic Inflammation Symposium  2012 

     More details

  • S100A8およびS100A9タンパク質の新規受容体の探索とそのがん進展における役割

    第71回日本癌学会学術総会  2012 

     More details

  • 細胞内導入型RAGE阻害ペプチドの開発

    日本生物工学会西日本支部創立30周年記念シンポジウム・講演会  2012 

     More details

  • C型ポツリヌス毒素変換ファージのゲノム情報を基盤とした偽溶原性メカニズムの解析

    第35回日本分子生物学会年会  2012 

     More details

  • TRAF6とSARM1によるPINK1のユビキチン化はマイトファジーの誘導に必須である

    第35回日本分子生物学会年会  2012 

     More details

  • RAGE高発現がもたらす細胞形態変化と細胞運動亢進の分子機構の解析

    第34回日本分子生物学会年会  2011 

     More details

  • S100タンパク質受容体の統合解析

    第34回日本分子生物学会年会  2011 

     More details

  • 多機能受容体RAGEの下流信号伝達機構の解明

    第70回日本癌学会学術総会  2011 

     More details

  • Mitochondral malfunction by PINK1 kn0ckdown augments apoptosis induced by adenoviirus carrying REIC/Dkk-3

    2011 

     More details

  • Promotion of migration and invasion of osteosarcoma cells by S100A7 through RAGE

    2011 

     More details

  • PINK1 knockdown increases the apoptosis induced by adenovirus-mediated REIC/Dkk-3

    日本組織培養学会第84回大会  2011 

     More details

  • REIC/Dkk-3 の扁平上皮における発現とその制御因子の探索

    日本組織培養学会第84回大会  2011 

     More details

  • Anti-proliferative effect of microRNA-34b/c adenoviral gene transfer on malignant pleural mesotheliomas

    2011 

     More details

  • TIRAP is a critical transducer of RAGE-mediated inflammatory signaling

    2011 

     More details

  • 膿疱性乾癬の病態解明とその対策に向けて - S100A8およびS100A9タンパク質の新規受容体の探索とその機能解析-

    稀少難治性皮膚疾患に関する調査研究班平成23年度第1回総会  2011 

     More details

  • PINK1発現抑制によるミトコンドリア機能不全はREIC/Dkk-3のアポトーソシス誘導効果を増強する

    第70回日本癌学会学術総会  2011 

     More details

  • S100A7による骨肉腫細胞の遊走・浸潤の多機能受容体RAGEを介した制御機構

    第70回日本癌学会学術総会  2011 

     More details

  • Mitochondrial dysfunction by PINK1 knockdown augments apotosis induced by a gene-therspeutic adenovirus carrying REIC/Dkk-3

    第34回日本分子生物学会年会  2011 

     More details

  • 正常皮膚におけるREIC/Dkk-3の発現制御因子の検索

    第34回日本分子生物学会年会  2011 

     More details

  • 悪性胸膜中皮腫に対するマイクロRNA34b/cを用いたアデノウイルス遺伝子治療の可能性

    第70回日本癌学会学術総会  2011 

     More details

  • Endocytotic internalization of Cy3-labeled REIC/Dkk-3 protein by differentiated ES cells and iPS cells

    2010 

     More details

  • KFL11とKLF15によるUCP1の発現制御

    第33回日本分子生物学会年会・第83回日本生化学会大会 合同大会  2010 

     More details

  • REIC/Dkk-3 gene therapy suppresses peritoneal dissemination of scirrhous gastric carcinoma

    2010 

     More details

  • TAE226, a bis-anilino pyrimidine compound, specifically inhibits the EGFR mutant kinase to show anti-tumor effect on non-small cell lung cancer with EGFR mutation

    2010 

     More details

  • Frequent silencing of tumor suppressive miR-34b/c by aberrant methylation in malignant pleural mesothelioma

    2010 

     More details