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

基本信息

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

项目摘要

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学基因的选择性表达应对肿瘤产生首选的细胞毒性作用，并为癌症治疗提供有用的策略。由于基因扩增和/或转录升高，C-ERBB-2和Midkine基因经常在人乳腺癌中过度表达。因此，我们检查了C-ERBB-2和中胺基因的启动子区域的可能用途，以更优选地表达乳腺癌细胞中的自杀基因。使用C-ERBB-2启动子区域的缺失突变体的本记者基因测定表明，251 bp（来自transcriplials启动站点的-213/+38），但不能直接转录相关的荧光素酶基因比SV40直接促进剂在术中乳腺癌的促进症更好。同样，中币基因的1.0 Kb启动子区域也可以激活人类乳腺癌细胞中融合的记者基因和自杀基因的转录。对比，251 bp片段介导的Pro…在非胸腺癌细胞中和正常成纤维细胞中更多的运动活性低于SV40启动子的活性。因此，126 bp片段（-213/-87）包含一个严格的元素，该元素负责乳腺癌细胞中首选的转录活性。电泳迁移率转移测定法表明，转录因子可能涉及肿瘤特异性。与含有与251 bp启动子（P256-TK）相关的质粒DNA的质粒DNA的交易，导致对乳腺癌的Ganciclovir（GCV）的敏感性提高，但在非肉类癌细胞中却没有提高。将GCV施用到带有人乳腺肿瘤的裸鼠中，这些小鼠被P256-TK DNA转移，这抑制了随后的移植肿瘤生长。这些结果表明，与这1.0 KB中心或251 BP C-ERBB-2启动子相关的自杀基因可能是针对乳腺癌特有的可行治疗策略。较少的