论文部分内容阅读
目的 探讨逆转录病毒载体介导单纯疱疹病毒Ⅰ型胸苷激酶 (HSV1 tk)结合环氧鸟苷(GCV) ,对雄激素非依赖性前列腺癌 (C4 2、PC3 )的治疗效果。 方法 通过DNA重组技术构建含tk基因的逆转录病毒载体 ,经转染包装细胞系PA3 17,选择出高病毒滴度的生产细胞系 (VPC) ,PCR、RT PCR检测VPC内HSV1 tk基因的整合和表达。VPC与癌细胞按不同比例混合培养 ,以硫基罗丹明B(SRB)法测定GCV治疗后癌细胞生长情况。 结果 经阳性克隆筛选 ,得到高滴度并能稳定表达HSV1 tk基因的VPC细胞。逆转录病毒载体介导的HSV1 tk基因对雄激素非依赖性前列腺癌C4 2、PC3均具有杀伤效应 ,光镜下没有发现凋亡小体 ;经比较 ,HSV1 tk基因对PC3的治疗效果最好 ,差别具有显著性意义。 结论 逆转录病毒载体介导的HSV1 tk基因对雄激素非依赖性前列腺癌细胞有杀伤作用 ,其机制是通过细胞毒作用
Objective To investigate the therapeutic effect of retroviral vector mediated HSV1 tk combined with guanosine monophosphate (GCV) on androgen independent prostate cancer (C4 2, PC3). Methods The recombinant retroviral vector containing tk gene was constructed by DNA recombination technique. After transfecting the packaging cell line PA3 17, a high virus titer production cell line (VPC) was selected. The integration of HSV1 tk gene in VPC was detected by PCR and RT PCR And expression. VPC and cancer cells were mixed at different ratios and the growth of cancer cells after GCV treatment was determined by the method of thio-rhodamine B (SRB). Results Positive clones were screened to obtain high titer VPC cells stably expressing HSV1 tk gene. HSV1 tk gene mediated by retroviral vector has anti-HSC1 and anti-prostate cancer C4 2 and PC3 all have killing effect, and no apoptotic body is found under light microscope; HSV1 tk gene has the best therapeutic effect on PC3 , The difference is significant. Conclusion The retroviral vector-mediated HSV1 tk gene has an anti-androgen-independent prostate cancer cell killing effect through the mechanism of cytotoxicity