低酸素微小環境における胃癌細胞の悪性形質獲得の機序解明と治療標的分子の探索

2014 Fiscal Year Annual Research Report

• Project/Area Number: 23390329
• Research Institution: Osaka City University
• Principal Investigator: 八代 正和 大阪市立大学, 医学(系)研究科(研究院), 准教授 (60305638)
• Project Period (FY): 2011-04-01 – 2015-03-31
• Keywords: スキルス胃癌 / 癌幹細胞 / 癌由来線維芽細胞 / 低酸素

Outline of Annual Research Achievements

癌の増殖進展により増加した癌組織低酸素領域は、癌細胞の悪性度を促進することが報告されている。しかし、この悪性度亢進機序は明らかにされていない。そこで、低酸素がSGCとCAFとの相互作用におよぼす影響を検討し、周囲低酸素環境におけるc-Met阻害剤やCXCR4阻害剤の有用性を検討した。
【方法】SGC細胞株(OCUM-2M, OCUM-2MD3,OCUM-12)とCAF(CaF-53)を通常酸素環境と低酸素環境にて共培養し、増殖能，遊走能や、増殖因子産生、及び増殖因子受容体発現を、細胞数算定やRT-PCR，Western blotにて比較検討した。また、CAFの産生するCXCL12 (SDF-1α）をELISA法で測定し比較検討した。さらに低酸素環境において、c-Met阻害剤やCXCR4阻害剤が癌細胞の増殖能や遊走能におよぼす影響を検討した。
【結果】SGCはCAFと共培養することで、通常酸素では増殖能は1.3倍、遊走能は1.4倍に増加したのに対し、低酸素環境ではさらに、増殖能が1.6倍、遊走能が２倍に増加した。低酸素環境は通常酸素に比し、SGC細胞のCXCR4発現が亢進し、CAFのCXCL12産生も2.5倍亢進した。siRNA によりHIF-1αを抑制することで、CXCR4発現は低下した。低酸素環境におけるCAFのSGC増殖促進作用はCXCR4阻害剤投与により抑制された。またCAFのSGC浸潤促進作用はc-Met阻害剤により抑制された。
【結論】低酸素環境においてCAFが産生するHGFやCXCL12が促進され、HIF1αを介してc-HGF/c-MetシグナルやCXCL12/CXCR4シグナルが増強されることが明らかになった。c-Met阻害剤やCXCR4阻害剤は癌周囲微小環境における新規分子標的治療剤であることが示唆された。

Research Progress Status

26年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

26年度が最終年度であるため、記入しない。

Research Products

• [Journal Article] Asporin activates coordinated invasion of scirrhous gastric cancer and cancer-associated fibroblasts. (2015)
  • Author(s): Satoyoshi R, Kuriyama S, Aiba N, Yashiro M, Tanaka M.
  • Journal Title: Oncogene. Volume: 34 Pages: 650-660
  • DOI: 10.1038/onc.2013.584.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Epigenetic silencing of miR-210 increases the proliferation of gastric epithelium during chronic Helicobacter pylori infection. (2014)
  • Author(s): Kiga K, Mimuro H, Suzuki M, Shinozaki A, Kobayashi T, Sanada T, Minsoo Kim M, Ogawa M, Watanabe Y, Kayo H, Fukuda Y, Iwai H, Yashiro M, Kirikae T, Fukayama M, Fukao T, and Sasakawa C.
  • Journal Title: Nature communications Volume: 5 Pages: 4497
  • DOI: 10.1038/ncomms5497.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Cancer-associated fibroblasts might sustain the stemness of scirrhous gastric cancer cells via transforming growth factor-beta signaling. (2014)
  • Author(s): Hasegawa T, Yashiro M, Nishii T, Matsuoka J, Fuyuhiro Y, Morisaki T, Fukuoka T, Shimizu K, Shimizu T, Miwa A, Hirakawa K.
  • Journal Title: Int J Cancer Volume: 134 Pages: 1785-1795
  • DOI: 10.1002/ijc.28520
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Lysyl oxidase-like 2 (LOXL2) from stromal fibroblasts stimulates the progression of gastric cancer. (2014)
  • Author(s): Kasashima H, Yashiro M, Kinoshita H, Fukuoka T, Morisaki T, Masuda G, Sakurai K, Kubo N, Ohira M, Hirakawa K
  • Journal Title: Cancer Lett. Volume: 354 Pages: 438-446
  • DOI: 10.1016/j.canlet.2014.08.014.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] MicroRNA-145 is a potential prognostic factor of scirrhous type gastric cancer. (2014)
  • Author(s): Naito Y, Yasuno K, Tagawa H, Sakamoto N, Oue N, Yashiro M, Sentani K, Goto K, Shinmei S, Oo HZ, Yanagihara K, Hirakawa K, Yasui W.
  • Journal Title: Oncology Reports Volume: 32 Pages: 1720-1726
  • DOI: 10.3892/or.2014.3333.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Comparative proteomics analysis of gastric cancer stem cells (2014)
  • Author(s): Morisaki T, Yashiro M, Kakehashi A, Inagaki A, Kinoshita H, Fukuoka T, Kasashima H, Masuda G, Sakurai K, Kubo N, Muguruma K, Ohira M, Wanibuchi H, Hirakawa K.
  • Journal Title: PLoS One Volume: 9 Pages: e110736.
  • DOI: 10.1371/journal.pone.0110736.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The niche component periostin is produced by cancer-associated fibroblasts, supporting growth of gastric cancer through ERK activation. (2014)
  • Author(s): Kikuchi Y, Kunita A, Iwata C, Komura D, Nishiyama T, Shimazu K, Takeshita K, Shibahara J, Kii I, Morishita Y, Yashiro M, Hirakawa K, Miyazono K, Kudo A, Fukayama M, Kashima TG.
  • Journal Title: Am J Pathol. Volume: 184 Pages: 859-870
  • DOI: 10.1016/j.ajpath.2013.11.012.
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Stromal fibroblasts mediate extracellular matrix remodeling and invasion of scirrhous gastric carcinoma cells. (2014)
  • Author(s): 11.Yamaguchi H, Yoshida N, Takanashi M, Ito Y, Fukami K, Yanagihara K, Yashiro M, Sakai R.
  • Journal Title: PLoS One Volume: 9 Pages: -
  • DOI: 10.1371/journal.pone.0085485.
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Bone marrow derived mesenchymal stem cells are associated with the tumor microenvironment of gastric tumor (2014)
  • Author(s): Kasashima H, Yashiro M, Masuda G, Morisaki T, Fukuoka T, Hasegawa T, Hirakawa K
  • Organizer: International Society for Stem Cell Research (ISSCR) 12th Annual Meeting
  • Place of Presentation: Vancouver (Canada)
  • Year and Date: 2014-06-18 – 2014-06-18
• [Presentation] The stemness of gastric cancer stem cells is sustained by TGFβ produced from cancer-associated fibroblasts (2014)
  • Author(s): Yashiro M, Hasegawa T, Fukuoka T, Kinoshita H, Morisaki T, Kasashima H, Masuda G, Kubo N, Hirakawa K
  • Organizer: American Association for Cancer Research 105th Annual Meeting 2014
  • Place of Presentation: San Diego (USA)
  • Year and Date: 2014-04-08 – 2014-04-08
• [Presentation] Effect of cancer associated fibroblast on gastric cancer cells under hypoxia (2014)
  • Author(s): Kinoshita H, Yashiro M, Masuda G, Kasashima H, Morisaki T, Fukuoka T, Shibutani M, Yamazoe S, Sakurai K, Nagahara H, Kimura K, Toyokawa T, Amano R, Kubo N, Tanaka H, Muguruma K, Ohtani H, Maeda K, Ohira M, Hirakawa K
  • Organizer: American Association for Cancer Research 105th Annual Meeting 2014
  • Place of Presentation: San Diego (USA)
  • Year and Date: 2014-04-08 – 2014-04-08
• [Presentation] The effect of PGE2 receptor inhibitor on the microenviroment of scirrhous gastric cancer (2014)
  • Author(s): Fukuoka T, Yashiro M, Takeda H, Maruyama T, Kasashima H, Masuda G, Kinoshita H, Morisaki T, Sakurai K, Shibutani M, Yamazoe S, Kimura K, Nagahara H, Toyokawa T, Amano R, Kubo N, Tanaka H, Muguruma K, Ohtani H, Maeda K, Ohira M, Hirakawa K
  • Organizer: American Association for Cancer Research 105th Annual Meeting 2014
  • Place of Presentation: San Diego (USA)
  • Year and Date: 2014-04-08 – 2014-04-08
• [Presentation] Role of lysyl oxidase and lysyl oxidase-like 2 in gastric carcinoma (2014)
  • Author(s): Kasashima H, Yashiro M, Go Y, Masuda G, Kinoshita H, Tokumoto M, Morisaki T, Fukuoka T, Toyokawa T, Kubo N, Tanaka H, Muguruma K, Ohira M, Hirakawa K
  • Organizer: American Association for Cancer Research 105th Annual Meeting 2014
  • Place of Presentation: San Diego (USA)
  • Year and Date: 2014-04-07 – 2014-04-07
• [Remarks] 大阪市立大学大学院 医学研究科 腫瘍外科学 腫瘍病態学講座
  • URL: http://www.med.osaka-cu.ac.jp/surgical-oncology/relation/geriatric_oncology.html