Suicide gene therapy for intractable gastrointestinal cancer with tumor specific promoters

具有肿瘤特异性启动子的自杀基因治疗顽固性胃肠癌

基本信息

批准号：
13670580
负责人：
KAWAMURA Kiyoko
金额：
$ 2.62万
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：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-13670580/
关键词：
Gene therapy Tumor promoter HSV-TK Reporter assay c-erbB-2 Survivin Midkine c-erbB-2

项目摘要

We examined the transcriptional regulatory regions of the midkine gene, which is expressed in a variety of tumors but scarcely in normal cells, with luciferase reporter assays. We demonstrated that the most potent transcriptional activity was obtained with the 2335-bp upstream region tested in hepatocellular carcinoma and with the 609-bp region in pancreatic tumors. The 1041- and the 335-bp but not the 70-bp region contained cis-acting elements for transcriptional activation in both types of tumor cells. Transfection of the herpes simplex virus-thymidine kinase (HSV-TK) gene fused with the midkine regulatory region in tumor cells increased their susceptibility to a prodrug ganciclovir. The regulatory regions of the c-erbB-2 gene, which is highly expressed in breast and gastric cancer, were also analyzed with the same reporter assays. We showed that the activity of the 251-bp region was the strongest in tumors among other genomic fragments deleted from 5'-side. Tumor cells transfected with the 251-bp region linked with the HSV-TK gene was more sensitive to ganciclovir in vitro and in vivo than parent cells. We also investigated the transcriptional control regions of the survivin gene, which is highly expressed in the G2/M phase. Deletion of the regulatory regions from the 5'-side showed that the 500-bp region was responsible for the preferential expression in tumor cells but not in normal cells.

我们检查了中酮基因的转录调节区域，该区域在多种肿瘤中表达，但在正常细胞中几乎没有荧光素酶报告基因分析。我们证明，最有效的转录活性是在肝细胞癌中测试的2335 bp上游区域以及胰腺肿瘤中的609 bp区域获得的。 1041-和335 bp，但不包含两种类型的肿瘤细胞中转录激活的顺式作用元素。与肿瘤细胞中的Midkine调节区域融合的单纯胸膜 - 胸腺苷激酶（HSV-TK）基因的转染使其对前药Ganciclovir的敏感性增加了。还用相同的记者测定法分析了在乳腺癌和胃癌中高度表达的C-ERBB-2基因的调节区域。我们表明，251 bp区域的活性是从5'侧删除的其他基因组片段中肿瘤中最强的。用与HSV-TK基因相关的251 bp区域转染的肿瘤细胞对Ganciclovir在体外和体内更敏感。我们还研究了Survivin基因的转录控制区域，该区域在G2/M期高度表达。从5'侧的调节区域缺失表明500 bp区域是肿瘤细胞中的优先表达，而不是正常细胞中的优先表达。