Suicide gene therapy for gastrointestinal tumors using tumor-specific promoters

使用肿瘤特异性启动子进行胃肠道肿瘤的自杀基因治疗

• 批准号: 11671298
• 负责人: TAGAWA Masatoshi
• 金额: $ 2.24万
• 依托单位: Chiba Cancer Center Research Institute
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-11671298/
• 关键词: Gene therapy; Promoter; Suicide gene; Transcriptional regulation; c-erbB-2; Midkine; VEGF; c-erbB2; エレクトロポレーション; 転写調節; 消化器がん; レトロウイルス

Tumor-specific gene expression is important for improved safely and enhanced efficacy of gene therapy for cancer. We then examined promoter regions of the midkine gene which was expressed in tumors but not in normal surrounding tissues or that of the c-erbB-2 and vascular endothelial growth factor gene whose expressions were predominantly found in the tumors of upper gastrointesinal tract and a number of solid tumors, respectively. Our results were as follows. (1)The midkine gene was expressed in 8 out of 14 human esophageal specimens and 14 out of 15 hepatocellular carcinoma specimnens, whereas none of non-tumorous regions of the same patients were negative for the expression. (2)Reporter assays using deletion mutants of the promoter region of the midkine gene showed that the 550-bp fragment was responsible for tumor-specific transcriptional activation. (3)The 250-bp fragment of 5'-upstream region of the c-erbB-2 gene strongly drove the transcription of the fused reporter gene in tumors but not in normal fibroblasts. (4)The 1.2-kb promoter region of the vascular endothelial growth factor gene had a cis-acting element for hypoxic responsiveness. (5)Forced expression of the herpes simplex virus-thymine kinase gene using these linked promoters conferred increased sensitivity of the transfected vells to ganciclovir. (6)The deoxycytidine kinase/ara-C and the uracil phosphoribosyl transferase/5-FU systems were useful for cell killing together with an appropriate promoter.

肿瘤特异性基因的表达对于提高肿瘤基因治疗的安全性和有效性具有重要意义。然后，我们检测了中期因子基因的启动子区域，该基因在肿瘤中表达，而在正常周围组织中不表达，以及c-erbB-2和血管内皮生长因子基因的启动子区，它们分别在上胃肠道肿瘤和一些实体肿瘤中表达。我们的研究结果如下。(1)中期因子基因在14例人食道标本中有8例表达，在15例肝细胞癌标本中有14例表达，而在同一病例的非肿瘤区均无表达。(2)利用中期因子基因启动子区域的缺失突变体进行的报告分析表明，550-bp的片段负责肿瘤特异性转录激活。(3)c-erbB-2基因5‘-上游区的250个碱基片段强烈驱动融合报告基因在肿瘤中的转录，而在正常成纤维细胞中不能。(4)血管内皮细胞生长因子基因的1.2 kb启动子区域具有缺氧反应的顺式作用元件。(5)利用这些连锁启动子强制表达单纯疱疹病毒胸腺苷酶基因，可增加细胞对更昔洛韦的敏感性。(6)脱氧胞苷酶/阿糖胞苷C和尿嘧啶磷酸核糖转移酶/5-FU系统在合适的启动子作用下对细胞有较好的杀伤作用。

项目成果

Yoshida, Y. , Tagawa, M. et al.: "Impaired tumorigenicity of human pancreatic cancer cells retrovirally transduced with interleukin-12 or interleukin-15 gene."Cancer Gene Ther. 7. 324-331 (2000)

Yoshida, Y.、Takawa, M. 等人：“用白细胞介素 12 或白细胞介素 15 基因逆转录病毒转导的人胰腺癌细胞的致瘤性受损。”Cancer Gene Ther。

Matsubara, H., Kimura, M., Sugaya, M., Koide, Y., Gunji, Y., Takegana, K., Asano, A., Ochiai, T., Isono, K., Sakiyama, S.and Tagawa, M.: "Expression of wild-type p53 gene confers increased sensitivity to radiation and chemotherapeutic agents in human esop

松原 H.、木村 M.、菅谷 M.、小出 Y.、群治 Y.、竹金 K.、浅野 A.、落合 T.、矶野 K.、崎山 S. 和

Miyauchi M.,Tagawa M.et al.: "Frequent expression of midkine gene in esophageal cancer suggests a potential usage of its promoter for suicide gene therapy"Japanese Journal of Cancer Research. 90. 469-475 (1999)

Miyauchi M.，Takawa M.等人：“食道癌中中期因子基因的频繁表达表明其启动子在自杀基因治疗中的潜在用途”日本癌症研究杂志。

Matsubara,H.,Tagawa,M. et al.: "Expression of wild-type p53 gene confers increased sensitivity to radiation and chemotherapeutic agents in human esophageal carcinoma cells"Int J Oncol. 14. 1081-1085 (1999)

松原 H.，田川 M.

Yoshida H.,Tagawa M.et al.: "Induced immunity by the expression of interleukin-2 or granulocyte macrophagecolony stimulating factor gene in murine neuroblastoma cells generated antitumor response for established tumors"Cancer Gene Ther.. 6. 395-401 (1999)

Yoshida H.、Takawa M.等人：“通过在小鼠神经母细胞瘤细胞中表达白介素-2 或粒细胞巨噬细胞集落刺激因子基因诱导免疫，对已建立的肿瘤产生抗肿瘤反应”Cancer Gene Ther.. 6. 395-401 (1999)