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.

我们通过荧光素酶报告基因检测检查了中期因子基因的转录调控区，该基因在多种肿瘤中表达，但在正常细胞中几乎不表达。我们证明，在肝细胞癌中测试的 2335 bp 上游区域和在胰腺肿瘤中测试的 609 bp 区域获得了最有效的转录活性。 1041 bp 和 335 bp 区域包含用于两种类型肿瘤细胞转录激活的顺式作用元件，但 70 bp 区域不包含。与肿瘤细胞中的中期因子调节区融合的单纯疱疹病毒胸苷激酶（HSV-TK）基因的转染增加了它们对前药更昔洛韦的敏感性。 c-erbB-2 基因的调控区域在乳腺癌和胃癌中高度表达，也使用相同的报告基因检测进行了分析。我们发现，在肿瘤中 5' 侧删除的其他基因组片段中，251 bp 区域的活性最强。转染有HSV-TK基因的251bp区域的肿瘤细胞在体外和体内比亲代细胞对更昔洛韦更敏感。我们还研究了生存素基因的转录控制区，该基因在 G2/M 期高表达。删除 5' 侧的调控区表明 500 bp 区域负责在肿瘤细胞中优先表达，但在正常细胞中则不然。