組織特異的 non-coding RNA が制御する癌の病態形成メカニズムの解析

2016 Fiscal Year Research-status Report

• Project/Area Number: 16K11496
• Research Institution: Fukuoka University
• Principal Investigator: 安河内 友世 (川久保友世) 福岡大学, 薬学部, 講師 (70507813)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: がん / 浸潤 / non-coding RNA

Outline of Annual Research Achievements

同一患者由来の転移能の異なるヒト口腔扁平上皮癌細胞株2種(非転移株SQUU-A/高転移株SQUU-B)を用いた実験から、転移能を規定する成分がSQUU-B細胞培養上清由来エクソソーム中に存在すること、また、その転移能はエクソソームを介して非転移株SQUU-Aに移行することを報告した。このことは、単一癌巣内のそれぞれの癌細胞が各々の個性をもつエクソソームを分泌しており、それらを介した複雑な細胞間クロストークによって癌微小環境が規定され、この複雑な環境が予後を規定していることを示唆していた。これらの内容については、J Oral Biosci 誌 (Kawakubo-Yasukochi et al., 58: 33-38, 2016; Morioka et al., 58: 180-184, 2016) にて報告した。また、各種国内学会、国際学会で発表し、高い評価を受けた。

加えて、平成28年度におけるその後の解析から、エクソソーム中の当該浸潤・転移規定因子がmiRNAであることが判明し、網羅的miRNA-mRNA統合解析を実施した結果、その標的候補miRNAを23個 (既知のmiRNA 4個 ＋ 未知のmiRNA 19個) に絞ることができた。また、上記23種のmiRNAのうち、どのmiRNAがヒト口腔扁平上皮癌の浸潤・転移能を規定しているのかを検討したところ、浸潤・転移を規定する数種のmiRNAの同定まで行うことができた。

Current Status of Research Progress

2: Research has progressed on the whole more than it was originally planned.

Reason

当初の研究計画通りの進捗状況である。

Strategy for Future Research Activity

平成28年度に得られた結果から、標的miRNAの浸潤・転移促進能が他の癌種（各種扁平上皮癌、腺癌）にも有効であるかマトリゲル浸潤アッセイにて検討を行う。in vivoにおける評価系では、転移規定miRNAの遺伝子導入を行った場合に、非転移株であるSQUU-Aがin vitro同様転移(浸潤)能を獲得するか、担癌ヌードマウスを用いた頸部リンパ節転移の評価で検討する。また、miRNA inhibitor libraryを用いて、標的miRNAに対する特異的阻害物質の探索を行い、in vitro、in vivoの評価系で浸潤・転移抑制効果を検討する。これらのことから、口腔扁平上皮癌転移規定因子としてのmiRNAおよびその阻害因子を同定し、in vitroおよびin vivoにおける機能評価を行うことで、当該miRNAや標的遺伝子のmiRNA結合部位配列の異常が、ヒト口腔扁平上皮癌予後予測因子として活用可能か否か方向づける。

Research Products

• [Journal Article] Differential roles of carboxylated and uncarboxylated osteocalcin in prostate cancer growth. (2016)
  • Author(s): Hayashi Y, Kawakubo-Yasukochi T, Mizokami A, Takeuchi H, Nakamura S, Hirata M.
  • Journal Title: J Cancer Volume: 7 Pages: 1605-1609
  • DOI: 10.7150/jca.15523
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Uncarboxylated osteocalcin increases serum nitric oxide levels and ameliorates hypercholesterolemia in mice fed an atherogenic diet. (2016)
  • Author(s): Kondo A, Kawakubo-Yasukochi T (corresponding author), Mizokami A, Chishaki S, Takeuchi H, Hirata M.
  • Journal Title: eJBio Volume: 13 Pages: 22-28
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Effect of collagen type I or human fibronectin on imatinib cytotoxicity in oral squamous cell carcinoma. (2016)
  • Author(s): Morioka M, Hazekawa M, Kawakubo-Yasukochi T, Nishinakagawa T, Nakamura S, Nakashima M.
  • Journal Title: Pharmacology & Pharmacy Volume: 7 Pages: 255-263
  • DOI: 10.4236/pp.2016.77032
  • Peer Reviewed / Open Access
• [Journal Article] Exosomes from oral squamous carcinoma cell lines, SQUU-A and SQUU-B, define the tropism of lymphatic dissemination. (2016)
  • Author(s): Morioka M, Kawakubo-Yasukochi T (corresponding author), Hayashi Y, Hazekawa M, Nishinakagawa T, Ono K, Kawano S, Nakamura S, Nakashima M.
  • Journal Title: J Oral Biosci Volume: 58 Pages: 180-184
  • DOI: 10.1016/j.job.2016.05.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Maternal oral administration of osteocalcin protects offspring from metabolic impairment in adulthood. (2016)
  • Author(s): Kawakubo-Yasukochi T, Kondo A, Mizokami A, Hayashi Y, Chishaki S, Nakamura S, Takeuchi H, Hirata M.
  • Journal Title: Obesity Volume: 24 Pages: 895-907
  • DOI: 10.1002/oby.21447
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Long-term oral administration of osteocalcin induces insulin resistance in male mice fed a high-fat, high-sucrose diet. (2016)
  • Author(s): Yasutake Y, Mizokami A, Kawakubo-Yasukochi T, Chishaki S, Takahashi I, Takeuchi H, Hirata M.
  • Journal Title: Am J Physiol Endocrinol Metab Volume: 310 Pages: E662-E675
  • DOI: 10.1152/ajpendo.00334.2015
  • Peer Reviewed / Open Access
• [Journal Article] The SQUU-B cell line spreads its metastatic properties to nonmetastatic clone SQUU-A from the same patient through exosomes. (2016)
  • Author(s): Kawakubo-Yasukochi T (corresponding author), Morioka M, Hayashi Y, Nishinakagawa T, Hazekawa M, Kawano S, Nakamura S, Nakashima M.
  • Journal Title: J Oral Biosci Volume: 58 Pages: 33-38
  • DOI: 10.1016/j.job.2015.10.001
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Hazekawa M, Morioka M, Nishinakagawa T, Kawakubo-Yasukochi T, Nakamura S, Nakashima M. (2016)
  • Author(s): Assessment of cytotoxicity of imatinib for oral squamous cell carcinoma by a real-time cell analysis system
  • Journal Title: eJBio Volume: 13 Pages: 56-62
  • Peer Reviewed / Open Access
• [Presentation] PBA投与による血中マウスDSS誘発性IBD発症抑制性物質の探索 (2017)
  • Author(s): 小野和彦、櫨川舞、西中川拓也、安河内（川久保）友世、中島学
  • Organizer: 日本薬学会第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-24 – 2017-03-27
• [Presentation] PBA代謝物PAGlnの抗炎症作用に関するin vitro評価 (2017)
  • Author(s): 櫨川舞、小野和彦、西中川拓也、安河内友世、中島学
  • Organizer: 日本薬学会第137年会
  • Place of Presentation: 仙台
  • Year and Date: 2017-03-24 – 2017-03-27
• [Presentation] The significance of BP180 expression in oral squamous carcinoma cell invasion (2017)
  • Author(s): Yasukochi A, Morioka M, Kawakubo-Yasukochi T, Nakashima M, Nakamura S.
  • Organizer: The 5th ASHNO Congress 2017, Congress of Asian Society of Head and Neck Oncology
  • Place of Presentation: Bali, Indonesia
  • Year and Date: 2017-03-23 – 2017-03-25
  • Int'l Joint Research
• [Presentation] Exosome-mediated crosstalk determines invasiveness and organotropic metastasis in oral squamous cell carcinoma (2017)
  • Author(s): Morioka M, Kawakubo-Yasukochi T, Hazekawa M, Nishinakagawa T, Ono K, Kawano S, Nakamura S, Nakashima M.
  • Organizer: The 5th ASHNO Congress 2017, Congress of Asian Society of Head and Neck Oncology
  • Place of Presentation: Bali, Indonesia
  • Year and Date: 2017-03-23 – 2017-03-25
  • Int'l Joint Research
• [Presentation] 卵巣がんにおけるsiRNA-PLGAハイブリッドミセル製剤の遺伝子抑制による治療効果 (2017)
  • Author(s): 櫨川舞、西中川拓也、安河内友世、中島学
  • Organizer: 第90回日本薬理学会年会
  • Place of Presentation: 長崎
  • Year and Date: 2017-03-15 – 2017-03-17
• [Presentation] Exosomes from oral squamous carcinoma cells define the tropism of invasiveness and lymphatic dissemination (2017)
  • Author(s): Morioka M, Kawakubo-Yasukochi T, Nakamura S, Nakashima M.
  • Organizer: Kyudai Oral Bioscience
  • Place of Presentation: Fukuoka, JAPAN
  • Year and Date: 2017-02-11
  • Int'l Joint Research
• [Presentation] Uncarboxylated osteocalcin prevents cancer progression through multiple mechanisms including immune system (2017)
  • Author(s): Hayashi Y, Kawakubo-Yasukochi T, Mizokami A, Nakamura S, Hirata M.
  • Organizer: Kyudai Oral Bioscience
  • Place of Presentation: Fukuoka, JAPAN
  • Year and Date: 2017-02-11
  • Int'l Joint Research
• [Presentation] Link of BP180 bullous pemphigoid autoantigen to invasion of oral squamous carcinoma cell (2016)
  • Author(s): Yasukochi A, Morioka M, Kawakubo-Yasukochi T, Obayashi K, Nakashima M, Nakamura S.
  • Organizer: Australian and New Zealand Head & neck cancer society
  • Place of Presentation: Auckland, New Zealand
  • Year and Date: 2016-10-25 – 2016-10-27
  • Int'l Joint Research
• [Presentation] Intercelluler crosstalk defined by exosomes from oral squamous carcinoma cells determines its pathological process (2016)
  • Author(s): Morioka M, Kawakubo-Yasukochi T, Hazekawa M, Nishinakagawa T, Ono K, Kawano S, Nakamura S, Nakashima M.
  • Organizer: Australian and New Zealand Head & neck cancer society
  • Place of Presentation: Auckland, New Zealand
  • Year and Date: 2016-10-25 – 2016-10-27
  • Int'l Joint Research
• [Presentation] Assessment of cytotoxicity of anticancer reagents for oral squamous cell carcinomas using a real-time cell monitoring analysis system (2016)
  • Author(s): Hazekawa M, Morioka M, Nishinakagawa T, Yasukochi T, Nakashima M.
  • Organizer: Australian and New Zealand Head & neck cancer society
  • Place of Presentation: Auckland, New Zealand
  • Year and Date: 2016-10-25 – 2016-10-27
  • Int'l Joint Research
• [Presentation] オステオカルシンを用いた次世代生活習慣病改善へのアプローチ (2016)
  • Author(s): 安河内（川久保）友世、近藤皓彦、溝上顕子、竹内弘、平田雅人
  • Organizer: 第37回日本肥満学会
  • Place of Presentation: 東京
  • Year and Date: 2016-10-07 – 2016-10-08
  • Invited
• [Presentation] Application of real-time cell monitoring device in evaluation the IC50 levels of anticancer reagents (2016)
  • Author(s): Hazekawa M, Morioka M, Nishinakagawa T, Yasukochi T, Nakashima M.
  • Organizer: 第75回日本癌学会学術大会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-06 – 2016-10-08
• [Presentation] Identify of the cytotoxic factor secreted by tumor cell line (2016)
  • Author(s): Nishinakagawa T, Hazekawa M, Yasukochi T, Nakashima M.
  • Organizer: 第75回日本癌学会学術大会
  • Place of Presentation: 横浜
  • Year and Date: 2016-10-06 – 2016-10-08
• [Presentation] 癌の増殖における非(低)カルボキシル化オステオカルシンの機能 (2016)
  • Author(s): 林慶和、川久保-安河内友世、溝上顕子、中村誠司、竹内弘、平田雅人
  • Organizer: 第58回歯科基礎医学会学術大会
  • Place of Presentation: 札幌
  • Year and Date: 2016-08-24 – 2016-08-26
• [Presentation] 口腔扁平上皮癌細胞由来エクソソームによる細胞間クロストークと病態形成 (2016)
  • Author(s): 森岡政彦、川久保-安河内友世、中村 誠司
  • Organizer: 第58回歯科基礎医学会学術大会
  • Place of Presentation: 札幌
  • Year and Date: 2016-08-24 – 2016-08-26
• [Presentation] The effect of uncarboxylated osteocalcin on cancer growth (2016)
  • Author(s): Hayashi Y, Kawakubo-Yasukochi T, Mizokami A, Nakamura S, Takeuchi H, Hirata M.
  • Organizer: The 9th Korea-Japan conference on cellular signaling for young scientists
  • Place of Presentation: Seol, Korea
  • Year and Date: 2016-07-21 – 2016-07-23
  • Int'l Joint Research
• [Presentation] The role of uncarboxylated osteocalcin in prevention of early atherosclerosis (2016)
  • Author(s): Kondo A, Kawakubo-Yasukochi T, Mizokami A, Takeuchi H, Hirata M.
  • Organizer: The 9th Korea-Japan conference on cellular signaling for young scientists
  • Place of Presentation: Seol, Korea
  • Year and Date: 2016-07-21 – 2016-07-23
  • Int'l Joint Research
• [Presentation] New functions of osteocalcin in cancer progression (2016)
  • Author(s): Hayashi Y, Kawakubo-Yasukochi T, Nakamura S, Hirata M.
  • Organizer: The 57th Congress of the Korean Association of Oral Maxillofacial Surgeons
  • Place of Presentation: Gunsan, Korea
  • Year and Date: 2016-04-22 – 2016-04-24
  • Int'l Joint Research