研究者詳細 - 阪口　政清

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

Masakiyo Sakaguchi, Nam-ho Huh

AMINO ACIDS 37 ( 1 ) 77 - 77 2009年7月

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 査読

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年5月

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

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年4月

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

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

JOURNAL OF GENE MEDICINE 10 ( 4 ) 454 - 454 2008年4月

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

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年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

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

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

Peptides 2004, Proceedings 1025 - 1026 2005年

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

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年10月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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 >29 days. These results indicate that the RFB; culture of OUMS-29/H-11 cells is useful for screening and developing new drugs.

Micromechanics of the damage-induced cellular microstructure in single crystal Ni-based superalloys

M. Sakaguchi, M. Okazaki

Acta Metallurgica Sinica (English Letters) 17 ( 4 ) 361 - 368 2004年8月

記述言語：英語掲載種別：研究論文（学術雑誌）

An analytical method to investigate the morphological evolution of the cellular microstructure is explored and proposed. The method is essentially based on the Eshelby's micromechanics theory, and it is extended so as to be applied for a material system containing inclusions with high volume fraction, by employing the average stress field approximation by Mori and Tanaka. The proposed method enables us to discuss a stable shape of precipitate in the material system, which must be influenced by many factors: e.g., volume fraction of precipitate
Young's modulus ratio and lattice misfit between matrix and precipitate
external stress field in multiaxial state
and heterogeneity of plastic strain between matrix and precipitate. A series of numerical calculations were summarized on stable shape maps. The application of the method to predict the γ′ rafting in superalloys during creep showed that the heterogeneity of plastic strain between matrix and precipitates may play a significant role in the shape stability of the precipitate. Furthermore, it was shown that the method was successfully applied to estimate the morphology of the cellular microstructure formed in CMSX-4 single crystal Ni-based superalloy.

Scopus

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

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

CANCER RESEARCH 64 ( 12 ) 4155 - 4161 2004年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

Differential expression of S100C in thyroid lesions 査読

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年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

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

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年3月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

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

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年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

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

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

CANCER LETTERS 201 ( 2 ) 195 - 201 2003年11月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

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

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年6月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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

Involvement of arachidonic acid in nonimmunologic production of superoxide in mast cells 査読

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年4月

記述言語：英語掲載種別：研究論文（学術雑誌）出版者・発行元：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