Faculty Profiles - OUCHIDA Mamoru

Identification of novel target proteins for miR-17-92 cluster by proteomic analysis

Hirotaka Kanzaki, Mamoru Ouchida, Sachio Ito, Seiji Tamaru, Hiroko Hanafusa, Kenji Shimizu

CANCER RESEARCH 69 2009.5

MET gene amplification or EGFR mutation activate MET in lung cancers untreated with EGFR tyrosine kinase inhibitors International journal

Takafumi Kubo, Hiromasa Yamamoto, William W. Lockwood, Ilse Valencia, Junichi Sob, Michael Peyton, Masaru Jida, Hiroki Otani, Tetsuva Fujii, Mamoru Ouchida, Nagio Takigawa, Katsuyuki Kiura, Kenji Shimizu, Hiroshi Date, John D. Minna, Marileila Varella-Garcia, Wan L. Lam, Adi F. Gazdar, Shinichi Toyooka

INTERNATIONAL JOURNAL OF CANCER 124 ( 8 ) 1778 - 1784 2009.4

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

We analyzed MET protein and copy number in NSCLC with or without EGFR mutations untreated with EGFR tyrosine kinase inhibitors (TKIs). MET copy number was examined in 28 NSCLC and 4 human bronchial epithelial cell lines (HBEC) and 100 primary tumors using quantitative real-time PCR. Positive results were confirmed by array comparative genomic hybridization and fluorescence in-situ hybridization. Total and phospho-MET protein expression was determined in 24 NSCLC and 2 HBEC cell lines using Western blot. EGFR mutations were examined for exon 19 deletions, T790M, and L858R. Knockdown of EGFR with siRNA was performed to examine the relation between EGFR and MET activation. High-level MET amplification was observed in 3 of 28 NSCLC cell lines and in 2 of 100 primary lung tumors that had not been treated with EGFR-TKIs. MET protein was highly expressed and phosphorylated in all the 3 cell lines with high MET amplification. In contrast, 6 NSCLC cell lines showed phosphoMET among 21 NSCLC cell lines without MET amplification (p = 0.042). Furthermore, those 6 cell lines harboring phospho-MET expression without MET amplification were all EGFR mutant (p = 0.0039). siRNA-mediated knockdown of EGFR abolished phospho-MET expression in examined 3 EGFR mutant cell lines of which MET gene copy number was not amplified. By contrast, phospho-MET expression in 2 cell lines with amplified MET gene was not down-regulated by knockdown of EGFR. Our results indicated that MET amplification was present in untreated NSCLC and EGFR mutation or MET amplification activated MET protein in NSCLC. (C) 2008 Wiley-Liss. Inc.

DOI： 10.1002/ijc.24150

Hyperthermia-induced seizure-susceptible rats: a novel model of febrile seizures

Tomoji Mashimo, Iori Ohmori, Mamoru Ouchida, Yukihiro Ohno, Toshiko Tsurumi, Takafumi Miki, Minoru Wakamori, Yasuo Mori, Tadao Serikawa

NEUROSCIENCE RESEARCH 65 S47 - S47 2009

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

Ca(v)2.1 DYSFUNCTION MAY BE A GENETIC MODIFIER OF SEVERE MYOCLONIC EPILEPSY IN INFANCY

Iori Ohmori, Mamoru Ouchida, Takafumi Miki, Nobuyoshi Mimaki, Shigeki Kiyonaka, Teiichi Nishiki, Kazuhito Tomizawa, Yasuo Mori, Hideki Matsui

JOURNAL OF PHYSIOLOGICAL SCIENCES 59 515 - 515 2009

A CACNB4 mutation shows that altered Ca(v)2.1 function may be a genetic modifier of severe myoclonic epilepsy in infancy International journal

Iori Ohmori, Mamoru Ouchida, Takafumi Miki, Nobuyoshi Mimaki, Shigeki Kiyonaka, Teiichi Nishiki, Kazuhito Tomizawa, Yasuo Mori, Hideki Matsui

NEUROBIOLOGY OF DISEASE 32 ( 3 ) 349 - 354 2008.12

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

Mutations of SCN1A, encoding the voltage-gated sodium channel alpha l subunit, represent the most frequent genetic cause of severe myoclonic epilepsy in infancy (SMEI). The purpose of this study was to determine if mutations in other seizure susceptibility genes are also present and could modify the disease severity. All coding exons of SCN1B, GABRG2, and CACNB4 genes were screened for mutations in 38 SCN1A-mutation-positive SMEI probands. We identified one proband who was heterozygous for a de novo SCN1A nonsense mutation (R568X) and another missense mutation (R468Q) of the CACNB4 gene. The latter mutation was inherited from his father who had a history of febrile seizures. An electrophysiological analysis of heterologous expression system exhibited that R468Q-CACNB4 showed greater Ba(2+) current density compared with the wild-type CACNB4. The greater Ca(v)2.1 currents caused by the R468Q-CACNB4 mutation may increase the neurotransmitter release in the excitatory neurons under the condition of insufficient inhibitory neurons caused primarily by the SCN1A mutation. (C) 2008 Elsevier Inc. All rights reserved.

DOI： 10.1016/j.nbd.2008.07.017

EGFR変異とUracil-Tegafurによる肺腺癌術後補助療法の関連性についての検討

PROTEOMICS-BASED ANALYSIS OF INVASION-RELATED PROTEINS IN MALIGNANT GLIOMAS

T. Maruo, T. Ichikawa, H. Kanzaki, M. Onishi, S. Inoue, K. Yoshida, H. Kambara, M. Ouchida, K. Shimizu, I. Date

NEURO-ONCOLOGY 10 ( 6 ) 1143 - 1143 2008.12

末久弘, 豊岡伸一, 堀田勝幸, 内田亜希子, 宗淳一, 藤原義朗, 松尾恵太郎, 大内田守, 高田穣, 木浦勝行, 伊達洋至

岡山医学会雑誌 120 ( 3 ) 265 - 269 2008.12

Language：Japanese Publisher：岡山医学会

完全切除された肺腺癌患者に補助療法としてUracil-Tegafur(UFT)を投与した場合における、EGFR変異の予後判定因子としての可能性を検討した。また、in vitro系のEGFR遺伝子導入細胞株における5-FUの効果も検討した。UFT投与群68例、手術のみの経過観察群119例であった。EGFR変異は187例中79例に認め、49例がエクソン19欠失変異、30例がエクソン21点突然変異であった。女性、非喫煙者に有意に変異が多かった。単変量解析では、UFT投与群が経過観察群よりも、EGFR変異群が野生群よりも予後がよい傾向であった。多変量解析では、術後補助療法が独立して有意に予後を延長していた。EGFR変異群では術後補助療法による予後延長の効果は認めなかった。EGFR変異型細胞株は5-FU低感受性を有することが示唆された。

Other Link： https://search.jamas.or.jp/index.php?module=Default&action=Link&pub_year=2008&ichushi_jid=J00175&link_issn=&doc_id=20081205250002&doc_link_id=10.4044%2Fjoma.120.265&url=https%3A%2F%2Fdoi.org%2F10.4044%2Fjoma.120.265&type=J-STAGE&icon=https%3A%2F%2Fjk04.jamas.or.jp%2Ficon%2F00007_3.gif

Novel SCN5A Mutations and SNP in Patients with Brugada Syndrome

Kazufumi Nakamura, Daiji Miura, Kaoru Kobayashi, Mamoru Ouchida, Kenji Shimizu, Iori Ohmori, Nobuhiro Nishii, Satoshi Nagase, Hiroshi Morita, Yoshiki Hata, Takefumi Oka, Kengo Kusano, Tohru Ohe

CIRCULATION 118 ( 18 ) S981 - S981 2008.10

Novel SCN5A Mutations and SNP in Patients with Brugada Syndrome

Kazufumi Nakamura, Daiji Miura, Kaoru Kobayashi, Mamoru Ouchida, Kenji Shimizu, Lori Ohmorl, Nobuhiro Nishii, Satoshi Nagase, Hiroshi Morita, Yoshiki Hata, Takefumi Oka, Kengo Kusano, Tohru Ohe

CIRCULATION 118 ( 18 ) S1485 - S1485 2008.10

The allelic distribution of a single nucleotide polymorphism in the PDCD5 gene locus of Japanese non-small cell lung cancer patients. Reviewed

Nanba K, Toyooka S, Soh J, Tsukuda K, Yamamoto H, Sakai A, Ouchida M, Kobayashi N, Matsuo K, Koide N, Kusaka K, Shimizu K, Date H

Molecular medicine reports 1 667 - 671 2008.9

Single nucleotide polymorphism in reuced folate carrier-1 gene and methyleneterathydrofolate reductase gene in patients with osteosarcoma

Y. Morimoto, A. Yoshida, M. Ouchida, T. Kunisada, K. Shimizu, T. Ozaki

EJC SUPPLEMENTS 6 ( 9 ) 117 - 118 2008.7

成人T細胞白血病/リンパ腫(ATLL)における特異的DNAメチル化と病態との関連

佐藤妃映, 岡剛史, 神農陽子, 鷲尾佳奈, 高田尚良, 守都敏晃, 大原信哉, 大内田守, 大島孝一, 谷本光音, 宇都宮與, 高橋聖之, 吉野正

日本リンパ網内系学会会誌 48 85 - 85 2008.5

A screening test for the prediction of Dravet syndrome before one year of age International journal

Junri Hattori, Mamoru Ouchida, Junko Ono, Susumu Miyake, Satoshi Maniwa, Nobuyoshi Mimaki, Yoko Ohtsuka, Iori Ohmori

EPILEPSIA 49 ( 4 ) 626 - 633 2008.4

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

Purpose: Our aim was to develop a screening test to predict Dravet syndrome before the first birthday based on the clinical characteristics of infants and the SCN1A mutation analysis.
Methods: Ninety-six patients who experienced febrile seizures before the age of one were enrolled. The patients were divided into two groups-the Dravet syndrome group (n = 46) and the non-Dravet syndrome group (n = 50). We compared the clinical characteristics before one year of age of the two groups. We analyzed all coding exons of the SCN1A gene by the direct sequencing method. Scores from 0 to 3 were assigned to each risk factor based on the odds ratio and p-value.
Results: An age of onset of febrile seizure <= 7 months, a total number of seizures >= 5, and prolonged seizures lasting more than 10 min. were regarded as significant risk factors for Dravet syndrome. Other factors highly predictive of this syndrome were hemiconvulsions, partial seizures, myoclonic seizures, and hot water-induced seizures. A total clinical score of six or above was the cutoff value indicating a high risk of Dravet syndrome. SCN1A missense and truncated mutations were detected significantly more often in the Dravet syndrome group than in the non-Dravet syndrome group.
Discussion: This simple screening test was designed to be used by general pediatricians. It could help to predict Dravet syndrome before one year of age. If the sum of the clinical risk score is >= 6, then the performance of an SCN1A mutation analysis is recommended.

DOI： 10.1111/j.1528-1167.2007.01475.x

Rasmussen encephalitis associated with SCN1A mutation International journal

Iori Ohmori, Mamoru Ouchida, Katsuhiro Kobayashi, Yoshimi Jitsumori, Takushi Inoue, Kenji Shimizu, Hideki Matsui, Yoko Ohtsuka, Yoshihiro Maegaki

EPILEPSIA 49 ( 3 ) 521 - 526 2008.3

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

Mutations in the SCN1A gene, encoding the neuronal voltage-gated sodium channel alpha 1 subunit, cause SMEI, GEFS+, and related epileptic syndromes. We herein report the R1575C-SCN1A mutation identified in a patient with Rasmussen encephalitis. R1575C were constructed in a recombinant human SCN1A and then heterologously expressed in HEK293 cells along with the human beta 1 and beta 2 sodium channel accessory subunits. Whole-cell patch-clamp recording was used to define biophysical properties. The R1575C channels exhibited increased channel availability and an increased persistent sodium current in comparison to the wild-type. These defects of electrophysiological properties can result in neuronal hyperexitability. The seizure susceptibility allele may influence the pathogenesis of Rasmussen encephalitis in this case.

DOI： 10.1111/j.1528-1167.2007.01411.x

The association between RAD18 Arg302Gln polymorphism and the risk of human non-small-cell lung cancer International journal

Hirotaka Kanzaki, Mamoru Ouchida, Hiroko Hanafusa, Hiromasa Yamamoto, Hiromitsu Suzuki, Masaaki Yano, Motoi Aoe, Kazue Imai, Hiroshi Date, Kei Nakachi, Kenji Shimizu

JOURNAL OF CANCER RESEARCH AND CLINICAL ONCOLOGY 134 ( 2 ) 211 - 217 2008.2

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

Purpose The repair enzyme RAD18 plays a key role in the post-replication repair process in various organisms from yeast to human, and the molecular function of the RAD18 protein has been elucidated. Single nucleotide polymorphism (SNP) of arginine (Arg, CGA) or glutamine (Gln, CAA) at codon 302 is known on RAD18; however, the association between the SNP and the risk of any human cancers including non-small-cell lung cancer (NSCLC) has not been reported. We therefore investigated the relationship between the polymorphism and the development and progression of human NSCLC.
Methods The study population included 159 patients with NSCLC and 200 healthy controls. The SNP was genotyped by polymerase chain reaction with the confronting two-pair primer (PCR-CTPP) assay. Genotype frequencies were compared between patients and controls, and the association of genotypes with clinicopathological parameters was also studied.
Results The Gln/Gln genotype was significantly more frequent in NSCLC patients (20.7%) than in healthy controls (11.5%)(P = 0.003). The increased risk was detected in NSCLC patients with the Gln/Gln genotype [Odds ratio (OR) = 2.63, 95% confidence interval (CI)=1.38-4.98]. As to the relationship of the SNP with clinicopathological parameters of NSCLC, significantly higher risks were detected in lung squamous cell carcinoma (LSC) (OR = 4.40, 95% CI = 1.60-12.1).
Conclusions Our results suggested that Gln/Gln genotype of the RAD18 SNP has the increased risk of NSCLC, especially of LSC. This is the first report to provide evidence for an association between the RAD18 Arg302Gln polymorphism and human NSCLC risk.

DOI： 10.1007/s00432-007-0272-3

Decreased expression of the SIN3A gene, a candidate tumor suppressor located at the prevalent allelic loss region 15q23 in non-small cell lung cancer International journal

Hiromitsu Suzuki, Mamoru Ouchida, Hiromasa Yamamoto, Masaaki Yano, Shinichi Toyooka, Motoi Aoe, Nobuyoshi Shimizu, Hiroshi Date, Kenji Shimizu

LUNG CANCER 59 ( 1 ) 24 - 31 2008.1

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

To identify the tumor suppressor genes (TSG) associated with non-small cell lung cancers (NSCLC), we performed the loss of heterozygosity (LOH) analysis in NSCLC samples from 66 patients. We focused on the novel hot spot region on 15q14-24 with eight polymorphic microsatellite markers. Frequent allelic toss was detected in 33 of 48 informative cases (69%) at D15S984 on 15q23. We defined the fine map on the region and identified the SlN3A gene as a candidate TSG. The SIN3A gene product is a component of the histone deacetylase (HDAC) complex and plays essential rotes in early embryonic development and the proliferation and survival. of a variety of cells through the repression of diverse signaling pathways. Our expression analysis revealed more frequent down-regulation of the SIN3A mRNA in 19 of 31 cases (61%) of NSCLCs in comparison to those of other flanking genes (16-42%), albeit the correlation of the decreased expression with the LOH did not attain statistic significance. These results suggest that the attenuated function of SIN3A due to a decreased level of expression may result in epigenetic de-regulation of growth-related genes through histone acetylation, which leads to the tumorigenesis of tung cancer cells. To our knowledge, this is the first evidence of the down-regulation of the SlN3A gene in human cancer. (c) 2007 Elsevier Ireland Ltd. All rights reserved.

DOI： 10.1016/j.lungcan.2007.08.002

Increased DOI-induced wet-dog shakes in adrenocorticotropic hormone-treated rats are not affected by chronic imipramine treatment: Possible involvement of enhanced 5-HT2A-receptor expression in the frontal cortex

Yoshihisa Kitamura, Kazuhiko Shibata, Kozue Akiyama, Shizue Kirnoto, Yoshika Fujitani, Kouhei Kitagawa, Hirotaka Kanzaki, Mamoru Ouchida, Kenji Shimizu, Hiromu Kawasaki, Toshiaki Sendo, Yutaka Gomita

JOURNAL OF PHARMACOLOGICAL SCIENCES 106 ( 1 ) 100 - 106 2008.1

Language：English Publishing type：Research paper (scientific journal) Publisher：JAPANESE PHARMACOLOGICAL SOC

We examined the influence of imipramine, a traditional tricyclic antidepressant, on the binding to serotonin (5-HT)(2) receptors and levels of 5-HT2A-receptor mRNA in the frontal cortex of rats treated with adrenocorticotropic hormone (ACTH). Chronic treatment with ACTH significantly increased the binding of [H-3]-ketanserin to 5-HT2 receptors and the expression of 5-HT2A-receptor mRNA in the frontal cortex. However, it did not alter the concentration of 5-HT or 5-hydroxyindole acetic acid. The effect of chronic ACTH treatment on 5-HT2 receptor and 5-HT2A-receptor mRNA levels was not altered by the chronic administration of imipramine. Also, imipramine did not affect the hyperfunction of 5-HT2A receptors caused by chronic ACTH treatment. These findings suggest that chronic treatment with ACTH acts to increase 5-HT2A-receptor synthesis through increased gene transcription, without modulating presynaptic serotonergic neurotransmission.

DOI： 10.1254/jphs.FP0071615

Single nucleotide polymorphism in the RAD18 gene and risk of colorectal cancer in the Japanese population International journal

Hirotaka Kanzaki, Mamoru Ouchida, Hiroko Hanafusa, Akiko Sakai, Hiromasa Yamamoto, Hiromitsu Suzuki, Masaaki Yano, Motoi Aoe, Kazue Imai, Hiroshi Date, Kei Nakachi, Kenji Shimizu

ONCOLOGY REPORTS 18 ( 5 ) 1171 - 1175 2007.11

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

The RAD18 gene, located on the human chromosome 3p24-p25, plays a crucial role in post-replication repair (PRR) in various organisms from yeast to humans. In the human RAD18 gene, one coding single nucleotide polymorphism (SNP) at codon 302, encoding either arginine (Arg, CGA) or glutamine (Gln, CAA), was reported. Although the molecular function of the RAD18 protein came to be elucidated, the association between the RAD18 Arg302Gln polymorphism and the risk of human cancer development was not examined. Therefore, we investigated the relationship between the polymorphism and the development of human primary colorectal cancer (CRC). The Arg302Gln polymorphism in 100 patients with CRC and 200 healthy controls were genotyped by the polymerase chain reaction with confronting two-pair primer (PCR-CTPP) assay. The Gln/Gln genotype was significantly more frequent in CRC (18.0%) than in the healthy controls (11.5%) (p=0.046). The increased risk was detected in CRC patients with the Gln/Gln genotype (Odds ratio [OR], 2.10; 95% confidence interval [CI], 1.00 to 4.40). When the relationship of the SNP with clinicopathological parameters of CRC was investigated, particularly in the well-differentiated grade and in the lymph node metastasis (NI) CRC patients, significantly higher risks were detected (OR, 7.00; 95% CI, 1.19-41.1 and OR, 3.71; 95% CI, 1.30-10.6, respectively). These results suggested that the RAD18 Arg302Gln polymorphism is associated with the risk of CRC. This report provides evidence for an association between the RAD18 Arg302Gln polymorphism and human CRC risk.