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Cyclin D1を分子標的とした治療不能消化器癌に対する遺伝子治療研究

Research Project

Project/Area Number 12770684
Research Category

Grant-in-Aid for Encouragement of Young Scientists (A)

Allocation TypeSingle-year Grants
Research Field Digestive surgery
Research InstitutionOsaka University

Principal Investigator

山本 浩文  大阪大学, 医学系研究科, 助手 (30322184)

Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥2,400,000 (Direct Cost: ¥2,400,000)
Fiscal Year 2001: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2000: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywordscyclin D1 / 遺伝子治療 / アデノウイルス / アンチセンス / 消化器癌 / 大腸癌 / 肝癌 / アデノウィルス / アンチャンス / 食道癌
Research Abstract

(目的)Antisense cyclin D1(CyD1)を用いた消化器癌遺伝子治療の臨床応用のためには,いろいろの遺伝子異常背景をもった多数の癌細胞株を用いた検定が必須である。
(検討項目と結果)
A.癌細胞のCyD1量,遺伝子異常背景についての検討
大腸癌細胞株5種類,肝癌細胞株4種類について,RT-PCR法でCyD1 mRNA発現を,western blot法で蛋白量を検討した。またp53,K-raS, APC, beta-cateninの遺伝子背景について検討した。
B.Adenovirus発現系の構築
Simplified adenovirus transfer system(PNAS;95:2509-2514,1998)を用いて,感染により各種細胞株にantisense CyD1 mRNAを発現させる系を確立する。(進行状況)pcDNA3-CyD1plasmidより,CyD1cDNAを切り出し,adenovirus shattle vectorにantisense方向にsubcloningした。次に,adenovirus backbone vector(Easy1)とelectroporation法により特殊大腸菌の中で融合させ,recombinant AS CyD1 vectorを得た。同様に,遺伝子の組み込まれていないMock vector,およびGFPを発現するコントロールplasmid, FP+antisense CyD1を発現するplasmidの作製が完了した。さらに293細胞によってウイルス液を濃縮・精製して,生物学的効果判定の実験をすすめている。現在のところantisense CyD1は腫瘍の血管新生を抑制することがわかり、そのメカニズムを調べている。

Report

(2 results)
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Miyata H, Yamamoto H, et al.: "Overexpression of CDC25B overrides radiation-induced G2-M arrest and results in increased apoptosis in esophageal cancer cells"Cancer Res. 61. 3188-3193 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Miyake Y, Yamamoto H, et al.: "Extensive micrometastases to lymph nodes as a marker for rapid recurrence of colorectal cancer : a study of lymphatic mapping"Clin Cancer Res. 7. 1350-1357 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Takamasa I, Yamamoto H, et al.: "Construction of Preferential cDNA Microarray Specialized for Human Colorectal Carcinoma : Molecular Sketch of Colorectai Cancer"Biochem Biophys Res Commun. 285. 1244-1249 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Nishioka K, Yamamoto H, et al.: "Clinical significance of CDC25A and CDC25B expression in squamous cell carcinomas of the oesophagus"Br J Cancer. 85. 412-421 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Hayashi N, Yamamoto H, et al.: "Differential expression of cyclooxygenase-2(COX-2)in human bile duct epithelial cells and bile duct neoplasm"Hepatology. 34. 638-650 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] Miyata H, et al: "Overexpression of CDC25B overrides radiation-induced G2/M arrest and results in increased apoptosis in esophageal cancer cells."Cancer Res,. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Miyake Y, et al: "Extended micrometastases of lymph node as a maker for rapid recurrence of colorectal cancer."Clin Cancer Res,. (in press). (2001)

    • Related Report
      2000 Annual Research Report
  • [Publications] Takemasa I, et al: "Overexpression of CDC25B phosphatase as a novel marker of poor prognosis of human colorectal carcinoma."Cancer Res,. 60. 3043-50 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Shamma A, et.al: "Up-regulation of cyclooxygenase-2 in squamous carcinogenesis of the esophagus."Clin Cancer Res,. 6. 1229-38 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Tsujie M, et.al: "Expression of tumor suppressor gene p16(INK4) products in primary gastric cancer."Oncology. 58. 126-36 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Eguchi H, et.al: "Augmentation of antitumor activity of 5-fluorouracil by interferon alpha is associated with up-regulation of p27Kip1 in human hepatocellular carcinoma cells."Clin Cancer Res,. 6. 2881-90 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Miyake Y, et.al: "Quantification of micrometastases in lymph nodes of colorectal cancer using real-time fluorescence polymerase chain reaction."Int J Oncol,. 16. 289-93 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ooka M, et.al: "Selection of mRNA markers for detection of lymph node micrometastases in breast cancer patients."Oncol Rep,. 7. 561-6 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Okami J, et al: "Genetic detection for micrometastasis in lymph node of biliary tract carcinoma."Clin Cancer Res,. 6. 2326-32 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Kondo M, et.al: "Expression of interferon・alpha/beta receptor in human hepatocellular carcinoma."Int J Oncol,. 17. 83-8 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Miyata H, et.al: "CDC25B and p53 are independently implicated in radiation sensitivity for human esophageal cancers."Clin Cancer Res,. 6. 4859-65 (2000)

    • Related Report
      2000 Annual Research Report

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Published: 2000-04-01   Modified: 2016-04-21  

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