SUICIDEAND IMMUNE GENE THERAPY USING TUMOR SPECIFIC PROMOTOR FOR BREAST CANCER

使用乳腺癌肿瘤特异性启动子的自杀和免疫基因治疗

• 批准号: 12671190
• 负责人: MATSUBARA Hisahiro
• 金额: $ 2.43万
• 依托单位: CHIBA UNIVERSITY (2001)Chiba Cancer Center (Research Institute) (2000)
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671190/
• 关键词: RREAST CANCER; SUICIDE GENE THERAPY; TUMOR SPECIFIC PROMOTER; midkine; c-erbB-2; HSV-tk; 腫瘍特異的プロモーター; c-erB2; 遺伝子治療; サイトカイン遺伝子; 自殺遺伝子

A selective expression of suicide or immune gene(s) in tumor cells should produce a preferential cytotoxic effect on tumors and a useful strategy for cancer treatment. The c-erbB-2 and midkine gene are frequently overexpressed in human breast cancers as a result of gene amplification and/or elevated transcription. We therefore examined a possible usage of promoter regions of the c-erbB-2 and midkine gene to express a suicide gene preferentially in breast cancer cells. The present reporter gene assays using deletion mutants of the c-erbB-2 promoter region demonstrated that the 251-bp (-213/+38 from the transcriplional start site) but not the 125-hp fragment (-87/+38) could direct transcription of the linked luciferase gene better than the SV40 immediate early promoter in breast cancer cells. Also the 1.0 kb promoter region of the midkine gene could activate transcription of a fused reporter gene and suicide gene in human breast cancer cells. In contrast, the 251-bp fragment-mediated pro … More moter activity in nonbreast cancer cells and in normal fibroblasts was lower than the activity by the SV40 promoter. The 126-bp fragment (-213/-87) thereby contains a exacting element(s) which is responsible for the preferential transcriptional activity in breast cancer cells. An electrophoretic mobility shift assay suggested that a possible modification of a transcriptional factor was involved in the tumor specificity. Transaction with the plasmid DNA containing the herpes simplex virus-thymidine kinase gene linked with the 251-bp promoter (p256-TK) resulted in increased sensitivity to ganciclovir (GCV) in breast cancer but not in nonbreast cancer cells. Administration of GCV into nude mice bearing human breast tumors that were transfecled with the p256-TK DNA suppressed subsequent growth of the transplanted tumors. These results suggest that delivery of a suicide gene linked with the these 1.0 kb midkine or 251 bp c-erbB-2 promoter can be a feasible therapeutic strategy specific to breast cancer. Less

肿瘤细胞中自杀或免疫基因的选择性表达应该对肿瘤产生优先的细胞毒性作用，并成为癌症治疗的有用策略。由于基因扩增和/或转录升高，c-erbB-2 和中期因子基因经常在人类乳腺癌中过度表达。因此，我们研究了c-erbB-2和中期因子基因的启动子区域在乳腺癌细胞中优先表达自杀基因的可能用途。使用c-erbB-2启动子区域的缺失突变体进行的报告基因分析表明，在乳腺癌细胞中，251-bp（来自转录起始位点的-213/+38）而不是125-hp片段（-87/+38）可以比SV40立即早期启动子更好地指导连接的荧光素酶基因的转录。此外，中期因子基因的 1.0 kb 启动子区域可以激活人类乳腺癌细胞中融合报告基因和自杀基因的转录。相比之下，非乳腺癌细胞和正常成纤维细胞中 251 bp 片段介导的促运动活性低于 SV40 启动子的活性。因此，126 bp 片段 (-213/-87) 包含负责在乳腺癌细胞中优先转录活性的精确元件。电泳迁移率变动分析表明转录因子的可能修饰与肿瘤特异性有关。与含有与 251 bp 启动子 (p256-TK) 连接的单纯疱疹病毒胸苷激酶基因的质粒 DNA 进行处理，导致乳腺癌细胞对更昔洛韦 (GCV) 的敏感性增加，但在非乳腺癌细胞中则不然。将GCV注射到转染了p256-TK DNA的携带人乳腺肿瘤的裸鼠体内，抑制了移植肿瘤的后续生长。这些结果表明，递送与这些 1.0 kb midkine 或 251 bp c-erbB-2 启动子连接的自杀基因可能是乳腺癌特异性的可行治疗策略。较少的

项目成果

Maeda T, et al.: "A minimum c-erbB-2 promoter-mediated expression of herpes simplex virus thymidine kinase gene confers selective cytotoxicity of human breast cancer cells to ganciclovir"Cancer Gene Therapy. 8・11. 890-896 (2001)

Maeda T 等人：“单纯疱疹病毒胸苷激酶基因的最小 c-erbB-2 启动子介导的表达赋予人乳腺癌细胞对更昔洛韦的选择性细胞毒性”癌症基因治疗 8·11。 ）

Matsubara,H.,Tagawa,M. et al: "Combinatory anti-tumor effects of electroporation-mediated chemotherapy and wild-type p53 gene transfer to human esophageal cancer cells."Int J Oncol. (in press). (2001)

松原 H.，田川 M.

Miyauchi, M. , Tagawa, M. et al.: "Expression of herpes simplex virus-thymidine kinase gene controlled by a promoter region of the midkine gene confers selective cytotoxicity to ganciclovir in human carcinoma cells."Int J Cancer. 91. 723-727 (2001)

Miyauchi, M.，Takawa, M.等人：“受中期因子基因启动子区域控制的单纯疱疹病毒胸苷激酶基因的表达赋予更昔洛韦在人类癌细胞中的选择性细胞毒性。”Int J Cancer。

Matsubara H, et al.: "A minimal promoter region of the c-erbB2 gene that can drive the expression of suicide gene preferentially in cancer cells"Cancer Gene Therapy. 8・Supp.2. S17 (2001)

Matsubara H 等人：“c-erbB2 基因的最小启动子区域可以优先在癌细胞中驱动自杀基因的表达”Cancer Gene Therapy 8·Supp.2 (2001)。

Yu L, et al.: "Identification of a minimal c-erbB-2 promoter region that mediates preferential expression of a linked foreign gene in human breast cancer cells"International Journal of Oncology. 20・3. 607-610 (2002)

Yu L等人：“介导人乳腺癌细胞中连锁外源基因优先表达的最小c-erbB-2启动子区域的鉴定”国际肿瘤学杂志20·3（2002）。