Analysis of DNA methylation and microRNA expression in hepatocellular carcinoma

2017 Fiscal Year Annual Research Report

• Project/Area Number: 15K09017
• Research Institution: Kyoto Prefectural University of Medicine
• Principal Investigator: 安居 幸一郎 京都府立医科大学, 医学(系)研究科(研究院), 准教授 (30323695)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: マイクロRNA / DNAメチル化

Outline of Annual Research Achievements

ヒトの癌の多くでは、癌抑制遺伝子のプロモーター領域のDNAメチル化が亢進していて、その結果、癌を抑制する働きを持ったタンパクの発現が抑制されている。

本研究の目的は、癌遺伝子に結合することでその発現を負に制御している腫瘍抑制性マイクロRNA（tumor suppressor miRNA）のプロモーター領域のDNAメチル化が亢進しており、そのため、発癌を促進する働きを持つタンパクの発現が亢進するという機構を肝細胞癌において明らかにすることである。

今年度までに、肝細胞癌の癌部と非癌部におけるDNAメチル化をアレイで網羅的に探索し、癌部で過剰なメチル化が生じているプロモーター領域のCpG部位を同定した。また、同じペアサンプルで、miRNAの発現量をアレイで網羅的に解析し、癌部で有意に発現が変化しているmiRNAを同定した。DNAメチル化とmiRNAの実験結果を統合的に解析し、さらに研究を進めたところ、以下の結果を得た。（1）miR-214とmiR-199aが癌部で最も有意に発現量が低下しているmiRNAである。(2) miR-214とmiR-199aは染色体1q24.3領域に隣接して座位しており、それらのhost geneであるDNM3OSのプロモーターにより転写制御を受けている。(3)癌部ではDNM3OSのプロモーター領域のDNAメチル化によってmiR-214とmiR-199aの発現が抑制されている。(4)網羅的mRNA発現解析、アルゴリズム解析、ルシフェラーゼ・アッセイとウエスタンブロット実験から、miR-214とmiR-199aが標的とする癌遺伝子としてPSMD10とMETをそれぞれ同定した。

Research Products

• [Journal Article] Oncogenic miR-96-5p inhibits apoptosis by targeting the caspase-9 gene in hepatocellular carcinoma (2018)
  • Author(s): Iwai Naoto、Yasui Kohichiroh、Tomie Akira、Gen Yasuyuki、Terasaki Kei、Kitaichi Tomoko、Soda Tomohiro、Yamada Nobuhisa、Dohi Osamu、Seko Yuya、Umemura Atsushi、Nishikawa Taichiro、Yamaguchi Kanji、Moriguchi Michihisa、Konishi Hideyuki、Naito Yuji、Itoh Yoshito
  • Journal Title: International Journal of Oncology Volume: 53 Pages: 237-245
  • DOI: 10.3892/ijo.2018.4369
  • Peer Reviewed
• [Journal Article] A multicenter Phase II study of sorafenib in Japanese patients with advanced hepatocellular carcinoma and Child Pugh A and B class (2018)
  • Author(s): Suzuki Eiichiro、Kaneko Shuichi、Okusaka Takuji、Ikeda Masafumi、Yamaguchi Kensei、Sugimoto Rie、Aramaki Takeshi、Asagi Akinori、Yasui Kohichiroh、Sano Keiji、Hosokawa Ayumu、Kato Naoya、Ishii Hiroshi、Sato Tosiya、Furuse Junji
  • Journal Title: Japanese Journal of Clinical Oncology Volume: 48 Pages: 317～321
  • DOI: 10.1093/jjco/hyy010
  • Peer Reviewed
• [Journal Article] Risk estimation model for nonalcoholic fatty liver disease in the Japanese using multiple genetic markers (2018)
  • Author(s): Kawaguchi T, Shima T, Mizuno M, Mitsumoto Y, Umemura A, Kanbara Y, Tanaka S, Sumida Y, Yasui K, Takahashi M, Matsuo K, Itoh Y, Tokushige K, Hashimoto E, Kiyosawa K, Kawaguchi M, Itoh H, Uto H, Komorizono Y, Shirabe K, Takami S, Takamura T, Kawanaka M, Yamada R, Matsuda F, Okanoue T.
  • Journal Title: PLOS ONE Volume: 13 Pages: e0185490
  • DOI: 10.1371/journal.pone.0185490
  • Peer Reviewed / Open Access
• [Journal Article] Combination of PNPLA3 and TLL1 polymorphism can predict advanced fibrosis in Japanese patients with nonalcoholic fatty liver disease (2018)
  • Author(s): Seko Y, Yamaguchi K, Mizuno N, Okuda K, Takemura M, Taketani H, Hara T, Umemura A, Nishikawa T, Moriguchi M, Yasui K, Kamaguchi M, Nishioji K, Mochizuki N, Kobayashi M, Mori K, Tanaka S, Matsuura K, Tanaka Y, Itoh Y.
  • Journal Title: Journal of Gastroenterology Volume: 53 Pages: 438～448
  • DOI: 10.1007/s00535-017-1372-8
  • Peer Reviewed
• [Journal Article] Real-world efficacy of daclatasvir and asunaprevir with respect to resistance-associated substitutions (2017)
  • Author(s): Fujii Hideki、Umemura Atsushi、Nishikawa Taichiro、Yamaguchi Kanji、Moriguchi Michihisa、Nakamura Hideki、Yasui Kohichiroh、Minami Masahito、Tanaka Saiyu、Ishikawa Hiroki、Kimura Hiroyuki、Takami Shiro、Nagao Yasuyuki、Shima Toshihide、Itoh Yoshito
  • Journal Title: World Journal of Hepatology Volume: 9 Pages: 1064～1064
  • DOI: 10.4254/wjh.v9.i25.1064
  • Peer Reviewed
• [Journal Article] Insulin resistance increases the risk of incident type 2 diabetes mellitus in patients with non-alcoholic fatty liver disease (2017)
  • Author(s): Seko Yuya、Sumida Yoshio、Tanaka Saiyu、Mori Kojiroh、Taketani Hiroyoshi、Ishiba Hiroshi、Hara Tasuku、Okajima Akira、Umemura Atsushi、Nishikawa Taichiro、Yamaguchi Kanji、Moriguchi Michihisa、Kanemasa Kazuyuki、Yasui Kohichiroh、Imai Shunsuke、Shimada Keiji、Itoh Yoshito
  • Journal Title: Hepatology Research Volume: 48 Pages: E42～E51
  • DOI: 10.1111/hepr.12925
  • Peer Reviewed
• [Journal Article] ASPP2 suppresses invasion and TGF-β1-induced epithelial?mesenchymal transition by inhibiting Smad7 degradation mediated by E3 ubiquitin ligase ITCH in gastric cancer (2017)
  • Author(s): Gen Yasuyuki、Yasui Kohichiroh、Kitaichi Tomoko、Iwai Naoto、Terasaki Kei、Dohi Osamu、Hashimoto Hikaru、Fukui Hayato、Inada Yutaka、Fukui Akifumi、Jo Masayasu、Moriguchi Michihisa、Nishikawa Taichiro、Umemura Atushi、Yamaguchi Kanji、Konishi Hiroyuki、Naito Yuji、Itoh Yoshito
  • Journal Title: Cancer Letters Volume: 398 Pages: 52～61
  • DOI: 10.1016/j.canlet.2017.04.002
  • Peer Reviewed
• [Journal Article] Loss of PAR-3 protein expression is associated with invasion, lymph node metastasis, and poor survival in esophageal squamous cell carcinoma (2017)
  • Author(s): Kitaichi Tomoko、Yasui Kohichiroh、Gen Yasuyuki、Dohi Osamu、Iwai Naoto、Tomie Akira、Yamada Nobuhisa、Terasaki Kei、Umemura Atsushi、Nishikawa Taichiro、Yamaguchi Kanji、Moriguchi Michihisa、Sumida Yoshio、Mitsuyoshi Hironori、Naito Yuji、Zen Yoh、Itoh Yoshito
  • Journal Title: Human Pathology Volume: 62 Pages: 134～140
  • DOI: 10.1016/j.humpath.2017.01.009
  • Peer Reviewed
• [Journal Article] Hepatic nucleotide binding oligomerization domain-like receptors pyrin domain-containing 3 inflammasomes are associated with the histologic severity of non-alcoholic fatty liver disease (2017)
  • Author(s): Mitsuyoshi Hironori、Yasui Kohichiroh、Hara Tasuku、Taketani Hiroyoshi、Ishiba Hiroshi、Okajima Akira、Seko Yuya、Umemura Atsushi、Nishikawa Taichiro、Yamaguchi Kanji、Moriguchi Michihisa、Minami Masahito、Itoh Yoshito
  • Journal Title: Hepatology Research Volume: 47 Pages: 1459～1468
  • DOI: 10.1111/hepr.12883
  • Peer Reviewed
• [Presentation] Epigenetic silencing of miR-214-3p is associated with resistance to p53-induced apoptosis through up-regulation of gankyrin expression in hepatocellular carcinoma. (2017)
  • Author(s): Yasui K, Iwai N, Terasaki K, Gen Y, Itoh Y.
  • Organizer: Annual Meeting 2017 of the American Association for Cancer Research
  • Int'l Joint Research
• [Presentation] SOX2 promotes cell proliferation of esophageal squamous cell carcinoma through the miR-17-92 cluster /PTEN/AKT/mTORC1 pathway. (2017)
  • Author(s): Terasaki K, Gen Y, Yasui K, Iwai N, Dohi O, Itoh Y.
  • Organizer: Annual Meeting 2017 of the American Association for Cancer Research
  • Int'l Joint Research
• [Presentation] miR-96-5p functions as an oncogenic miRNA by inhibiting apoptosis in hepatocellular carcinoma. (2017)
  • Author(s): 2.Iwai N, Yasui K, Tomie A, Terasaki K, Kitaichi T, Dohi O, Gen Y, Itoh Y.
  • Organizer: Annual Meeting 2017 of the American Association for Cancer Research
  • Int'l Joint Research