2000 Fiscal Year Annual Research Report

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-基礎実験から臨床応用へ-

Research Project

Project/Area Number	12770684
Research Institution	Osaka University
Principal Investigator	山本浩文大阪大学, 医学系研究科, 助手 (30322184)
Keywords	cyclin D1 / 遺伝子治療 / アデノウィルス / アンチャンス / 消化器癌 / 大腸癌 / 肝癌 / 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-CyD1 plasmid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,GFP+antisense CyD1を発現するplasmidの作製が完了した。今後293細胞によってウイルス液を得て,生物学的効果判定の実験をすすめる。Antisense CyD1の生物学的効果検討としては,以下の項目を予定している。血管新生,マトリックス分解,アポトーシス,細胞周期,接着因子, 運動・浸潤能,造腫瘍性

Research Products

(11 results)

All Other

All Publications (11 results)

[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)
[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)
[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)
[Publications] Shamma A, et.al: "Up-regulation of cyclooxygenase-2 in squamous carcinogenesis of the esophagus."Clin Cancer Res,. 6. 1229-38 (2000)
[Publications] Tsujie M, et.al: "Expression of tumor suppressor gene p16(INK4) products in primary gastric cancer."Oncology. 58. 126-36 (2000)
[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)
[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)
[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)
[Publications] Okami J, et al: "Genetic detection for micrometastasis in lymph node of biliary tract carcinoma."Clin Cancer Res,. 6. 2326-32 (2000)
[Publications] Kondo M, et.al: "Expression of interferon・alpha/beta receptor in human hepatocellular carcinoma."Int J Oncol,. 17. 83-8 (2000)
[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)

2000 Fiscal Year Annual Research Report

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-基礎実験から臨床応用へ-

Principal Investigator

山本 浩文 大阪大学, 医学系研究科, 助手 (30322184)

Research Products

[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)

[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)

[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)

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

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

[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)

[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)

[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)

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

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

[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)

山本浩文大阪大学, 医学系研究科, 助手 (30322184)