Mouse mammary carcinoma cell lines release microvesicles containing precurcer VEGF-C

• Project/Area Number: 25462003
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: General surgery
• Research Institution: Osaka Medical College
• Principal Investigator: Ito Yuko 大阪医科大学, 医学部, 准教授 (40148432)
• Co-Investigator(Kenkyū-buntansha): Shibata Masa-aki 大阪保健医療大学, 保健医療学部, 教授 (10319543) ; OTSUKI Yoshinori 大阪医科大学, 医学部, 教授 (50140166)
• Co-Investigator(Renkei-kenkyūsha): AKAO Yukihiro 岐阜大学, 連合創薬医療情報研究科, 教授 (00222505) ; NABIL Eid 大阪医科大学, 医学部, 講師 (50570165)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥4,290,000 (Direct Cost: ¥3,300,000、Indirect Cost: ¥990,000); Fiscal Year 2015: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2014: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2013: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: microvesicle / exosome / VEGF-C / マウス乳癌細胞 / リンパ行性転移 / リンパ管新生 / オートクライン / microvesicles / 乳癌 / 乳癌のリンパ行性転移 / 転移性の差異 / microRNA / 低酸素 / exosomes / 乳癌細胞 / リンパ管形成 / matureVEGF-C / precursorVEGF-C / lymphangiogenesis / metastasis

Outline of Final Research Achievements

Using two mouse mammary carcinoma cell lines, Western blot analysis of VEGF-C demonstrated exosomal expression of premature VEGF-C (pre-VEGF-C) in both cell lines. To study target cells of that pre-VEGF-C, we used mouse endothelial cell line (UV2) which express VEGFR3. MVs in the cell lines enhanced cell proliferation and tube formation in MV-treated UV2 cells. To confirm VEGF-C/VEGFR3 interaction, Western blot and immunofluorescent analysis were performed for the MV-treated UV2 cells. After treated by MVs, UV2 strongly expressed phospho-Akt. Furthermore, neutrizing of VEGFR3 inhibited proliferation of UV2, whereas UV2 were treated by MVs. These results indicated that there was no significant difference of pre-VEGF-C between these MVs shed from low- and high-metastatic tumors, while both MVs enhanced lymphangiogenesis of endothelial cells. Highly metastatic BJMC3879 cell was suggested as another target cell of pre-VEGF-C packed in MVs, because this cell expressed VEGFR3.

Research Products

• [Journal Article] Indomethacin suppresses LAMP-2 expression and induces lipophagy and lipoapoptosis in rat enterocytes via the ER stress pathway. (2015)
  • Author(s): Narabayashi K, Ito Y, Eid N, Maemura K, Inoue T, Takeuchi T, Otsuki Y, Higuchi K
  • Journal Title: J Gastroenterol. Volume: 50 Issue: 5 Pages: 541-554
  • DOI: 10.1007/s00535-014-0995-2
  • Peer Reviewed / Open Access
• [Journal Article] Ethanol-induced damage in rat testis: oxidative stress-mediated germ cell apoptosis and vacuolization in Sertoli cells. (2015)
  • Author(s): Eid N, Ito Y, Otsuki Y.
  • Journal Title: Andrologia Volume: 47 Issue: 1 Pages: 3-3
  • DOI: 10.1111/and.12385
  • Peer Reviewed / Open Access
• [Journal Article] Anti-cancer fatty-acid derivative induces autophagic cell death through modulation of PKM isoforms expression profile mediated by bcr-abl in chronic myeloid leukemia. (2015)
  • Author(s): Shinohara H, Taniguchi K, Kumazaki M, Yamada N, Ito Y, Otsuki Y, Uno B, Hayakawa F, Minami Y, Naoe T, Akao Y.
  • Journal Title: Cancer Letters Volume: 360 Issue: 1 Pages: 28-38
  • DOI: 10.1016/j.canlet.2015.01.039
  • Peer Reviewed / Open Access
• [Journal Article] Organ-specific PTB1-associated microRNAs determine expression of pyruvate kinase isoforms. (2015)
  • Author(s): Taniguchi K, Ito Y, Sugito N, Kumazaki M, Shinohara H, Yamada N, Nakagawa Y, Sugiyama T, Futamura M, Otsuki Y, Yoshida K, Uchiyama K, Akao Y.
  • Journal Title: Sci Rep. Volume: 2 Issue: 1 Pages: 8647-8647
  • DOI: 10.1038/srep08647
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] MicroRNA-124 inhibits cancer cell growth through PTB1/PKM1/PKM2 feedback cascade in colorectal cancer. (2015)
  • Author(s): Taniguchi K, Sugito N, Kumazaki M, Shinohara H, Yamada N, Nakagawa Y, Ito Y, Otsuki Y, Uno B, Uchiyama K, Akao Y.
  • Journal Title: Cancer Lett. Volume: 1 Issue: 1 Pages: 17-27
  • DOI: 10.1016/j.canlet.2015.03.026
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Ethanol-induced hepatic autophagy: Friend or foe? (2015)
  • Author(s): Eid N, Ito Y, Otsuki Y.
  • Journal Title: World J Hepatol. Volume: 7 Issue: 9 Pages: 1154-1156
  • DOI: 10.4254/wjh.v7.i9.1154
  • Peer Reviewed / Open Access
• [Journal Article] Positive feedback of DDX6/c-Myc/PTB1 regulated by miR-124 contributes to maintenance of the Warburg effect in colon cancer cells. (2015)
  • Author(s): Taniguchi K, Sugito N, Kumazaki M, Shinohara H, Yamada N, Matsuhashi N, Futamura M, Ito Y, Otsuki Y, Yoshida K, Uchiyama K, Akao Y.
  • Journal Title: Biochim Biophys Acta. Volume: 9 Issue: 9 Pages: 1971-80
  • DOI: 10.1016/j.bbadis.2015.06.022
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] Mitophagy in steatotic hepatocytes of ethanol-treated wild-type and Parkin knockout mice (2015)
  • Author(s): Eid N, Ito Y, Otsuki Y
  • Journal Title: Am J Physiol Gastrointest Liver Physiol Volume: 309 Issue: 6 Pages: G513-G514
  • DOI: 10.1152/ajpgi.00254.2015
  • Peer Reviewed / Open Access
• [Journal Article] 3-DECENOIC ACID DERIVATIVES INDUCE AUTOPHAGIC CELL DEATH THROUGH TH EDOWN-REGULATION OF E2F1 (2013)
  • Author(s): SHINOHARA H.1,2, NOGUCHI S.2, KUMAZAKI M.2,3, YAMADA N.2, ITO T.1, OYAMA M., ITO Y., OTSUKI Y.,NAITO S., IINUMA M. AND AKAO Y.
  • Journal Title: Journal of Toxicology Research Volume: 3 Pages: 29-34
  • Peer Reviewed
• [Presentation] Ethanol enhances apoptosis, mitophagy and mitochondrial spheroid. (2016)
  • Author(s): Nabil Eid, Yuko Ito, Yoshinori Otsuki
  • Organizer: 第１２１回解剖学会
  • Place of Presentation: ビッグパッレット福島
  • Year and Date: 2016-03-29
• [Presentation] 高転移性マウス乳癌細胞はprecursor VEGF-Cをautocrineする (2016)
  • Author(s): 伊藤裕子、ナビル・イード、前村憲太朗、柴田雅朗、大槻勝紀
  • Organizer: 第１２１回解剖学会
  • Place of Presentation: ビッグパッレット福島
  • Year and Date: 2016-03-28
• [Presentation] Ethanol induces thymocyte apoptosis and activates autophagy in thymic macrophages. (2016)
  • Author(s): Tubasa Betsuyaku, Nabil Eid, Yuko Ito, Yoshihisa Tanaka, Kentaro Maemura, Yoshinori Otsuki
  • Organizer: 第１２１回解剖学会
  • Place of Presentation: ビッグパッレット福島
  • Year and Date: 2016-03-28
• [Presentation] マウス乳癌細胞はVEGF-C前駆体を内包したmicrovesiclesを分泌する (2015)
  • Author(s): 伊藤裕子、柴田雅朗、前村憲太朗、谷口高平、赤尾幸博、大槻勝紀
  • Organizer: 第７４回日本癌学会
  • Place of Presentation: 名古屋国際会議場
  • Year and Date: 2015-10-10
• [Presentation] Microvesicles released from two mouse mammary carcinoma cell lines contained premature VEGF-C (2015)
  • Author(s): Ito Y , Nabil Eid , Shibata M, Otsuki Y
  • Organizer: The 120th Annual Meeting of the Japanese Association of Anatomists
  • Place of Presentation: 神戸国際国際会議場
  • Year and Date: 2015-03-21 – 2015-03-23
• [Presentation] マウス乳癌細胞はprecursor VEGF-C を内包したexosomeを放出する (2014)
  • Author(s): 伊藤裕子、ナビル・イード、柴田雅朗、赤尾幸博、大槻勝紀
  • Organizer: 第119回日本解剖学会総会
  • Place of Presentation: 自治医科大学キャンパス
• [Remarks] 解剖学教室研究業績
  • URL: http://www.osaka-med.ac.jp/deps/an1/results.html
• [Remarks] 大阪医科大学研究支援センター 研究機構年報
  • URL: http://www.osaka-med.ac.jp/deps/kik/index.htm#hidden02