Clinical research of gene therapy against digestive tract cancers by using ras suppressor mutant.

ras抑制突变体抗消化道肿瘤基因治疗的临床研究

• 批准号: 07557086
• 负责人: KUZUMAKI Noboru
• 金额: $ 6.46万
• 依托单位: Hokkaido University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07557086/
• 关键词: ras gene; mutant; pancreatic cancer; esophageal cancer; retrovirus; adenovirus; gene therapy; 抑制変異体; ヒト食道癌; アデノウイルスベクター; ヒト膵癌; レトロウイルスベクター; すい臓癌

Our previous studies demonstrated that introduction of a dominant negative H-ras mutant, N116Y,inhibits the growth of various types of cancer cells in vitro. In this study, we testedthe efficacy of N116Y in blocking the growth of human pancreatic and esophageal cancer cells using viral vectors.We transfected an retrovirus-expression vector of N116Y,pZIP-NI16Y,into eight human pancreatic cancer cell lines with K-ras mutations. The growth of the pancreatic cancer cell lines was strongly inhibited, and the N116Y-expressing clones became less spread and lost their anchorage-independent growth ability. Furthermore, they were non-tumorigenic in vivo. Infection with Nl16Y adenovirus (AdCMV-N116Y) inhibitedthe in vitro growth of all esophageal cancer cell lines studied. In the AdCMV-N116Y virus-infected cells, progression into S phase was clearly blocked, and did not show the activation of Erk2 after EGF stimulation. Most importantly, direct injection of AdCMV-N116Y into the esophageal tumors in nude mice suppressed their growth significantly.These observations suggest that N116Y suppresses growth of human pancreatic and esophageal cancer cells in vitro and in vivo through the inhibition of Erk2 activation, and that N116Y is a potential candidate gene for gene therapy against human pancreatic and esophageal cancers.

我们以前的研究表明，引入显性负性H-ras突变体N116 Y，在体外抑制各种类型的癌细胞的生长。本研究利用逆转录病毒载体pZIP-NI 16 Y转染8株K-ras突变的人胰腺癌细胞系，观察N116 Y对人胰腺癌和食管癌细胞生长的抑制作用。胰腺癌细胞系的生长受到强烈抑制，N116 Y表达克隆变得不那么扩散，失去了它们的锚定非依赖性生长能力。此外，它们在体内无致瘤性。N116 Y腺病毒（AdCMV-N116 Y）感染食管癌细胞后，对所有食管癌细胞株的体外生长均有抑制作用。在AdCMV-N116 Y病毒感染的细胞中，进入S期的进程被明显阻断，并且在EGF刺激后没有显示Erk 2的活化。结果表明，N116 Y通过抑制Erk 2的活化，在体内外抑制人胰腺癌和食管癌细胞的生长，N116 Y是一个潜在的胰腺癌和食管癌基因治疗的候选基因。

项目成果

Ishihara, H ほか: "Specific detection of the precursor of ras p21 with a mouse monoclonal anti-C-terminal peptide antibody, SARA-K1" Journal of Immunological Methods. 185. 217-223 (1995)

Ishihara, H 等人：“用小鼠单克隆抗 C 端肽抗体 SARA-K1 特异性检测 ras p21 前体”，免疫学方法杂志 185. 217-223 (1995)。

Ishihara, H.et al: "Specific detection of the precursor of ras p21 with a mouse monoclonal anti-C-terminal peptide antibody, SARA-Kl." J.Immunol.Method. 185. 217-223 (1995)

Ishihara, H.等人：“用小鼠单克隆抗 C 端肽抗体 SARA-K1 特异性检测 ras p21 前体。”

Shichinohe, T.et al: "Suppression of pancreatic cancer by the dominant negative ras mutant, N116Y." J.of Surgical Research. 66. 125-130 (1996)

Shichinohe, T.等人：“显性失活 ras 突变体 N116Y 对胰腺癌的抑制。”

Yabunaka,N.et al: "Involvement of ras in the expression of glycolipid sulfotransferase in human renal cancer cells." Int.J.Cancer. 71. 620-623 (1997)

Yabunaka,N.et al：“ras 参与人肾癌细胞中糖脂磺基转移酶的表达。”

Shichinohe, T. et al: "Suppression of pancreatic cancer by the dominant negative ras mutant, N16Y" J. of Surgical Research. 66. 125-130 (1996)

Shichinohe, T. 等人：“显性失活 ras 突变体 N16Y 对胰腺癌的抑制”J. of Surgical Research。