HSV-tk/GCV系统对鼠膀胱癌的自杀基因治疗研究

Suicide gene therapy for mouse bladder cancer using HSV-tk/GCV system

目的:在体外和体内水平建立膀胱癌的单纯疱疹病毒-胸苷激酶/丙氧鸟苷(herpes simplex virus type thymidine Kinase/ganciclovir,HSV-tk/GCV)自杀基因系统,并验证该系统的有效性.方法:用逆转录病毒载体HSV-TK转染鼠膀胱癌细胞株T739,体外检测其对GCV的敏感性,在同基因鼠,分别建立T739和T739-TK膀胱癌腹腔肿瘤模型.当B超显示肿瘤直径约0.5～0.8cm时,腹腔GCV给药6天,然后观察肿瘤大小变化及动物存活时间.结果:GCV对T739-TK细胞有显著杀伤作用,而对T739细胞的生长无影响,成功的建立了T739同基因鼠膀胱癌腹腔肿瘤模型,体内实验得到相应结果,转染了TK基因的T739鼠肿瘤明显变小,甚至消失,而未转染TK基因的T739鼠肿瘤继续增大,GCV对其无明显治疗作用,转染了TK基因的T739鼠存活时间显著延长.结论:在体外和体内水平,表达TK基因的肿瘤细胞均被GCV有效杀伤,这表明HSV-tk/GCV系统有可能成为基因治疗膀胱癌的有效方法.

Objective ：To establish a suicide gene therapy system HSV- tk/C, CV for bladder cancer,a treatment in vitro as well as in vivo and to test its efficacy. Methods：Mouse bladder cancer cell line （T739） was transfected with retrovira[ vector HSV- tk gene.The sensitivity of T739 TK cells to C, CV was detected in vitro. In the mouse model of bladder tumor, T739 or T739 -TK was implanted beneath the peritoneum of syngeneic mice. When tumors grew to the size of 0.5 -- 0.8cm, intraperitoneal administration of GCV was carried out for 6 days. Changes of tumor size and survival rate of mice were observed. Results ： RT - PCR showed that TK gene was transferred into the T739 cell and expressed successfully. In vitro, T739 - TK cells became sensitive to low concentrations of GCV. In vivo studies showed similar re_suit. Significant tumor inhibition was found in the T739 - TK group after administration of GCV, and the survival time of mice was prolonged. Conclusion ：Tumor cells expressing HSV- tk gene were eradicated by administration of GCV in vitro as well as in vivo.