【摘要】 目的 :观察单纯疱疹病毒胸苷激酶 (HSV TK)丙氧鸟苷 (GCV)自杀基因系统对人宫颈癌Hela细胞系体外杀伤作用及其旁观效应。　方法 :采用脂质体转染法将GINaTK载体转入包装细胞PA317。取病毒上清液感染Hela细胞 ,得到带有HSV TK基因的Hela/TK细胞 ,并将其用于体外实验。　结果 :载体HSV TK导入了PA317细胞。体外实验结果显示 ,当Hela/TK细胞数占混合细胞 10 %时 ,低浓度 (10mg/L)的GCV就可将 5 0 %左右的肿瘤细胞杀死。GCV作用后的实验组Hela /TK细胞基因组经琼脂糖凝胶电泳后可观察到典型的DNA梯状条带。　结论 :逆转录病毒可介导HSV TK基因转入人宫颈癌Hela细胞并获稳定表达 ,HSV TK/GCV自杀基因系统在体外对宫颈癌细胞有杀伤作用 ,且存在明显的旁观效应。更多还原

【Abstract】 Objective:To investigate the killing effect and the bystander effect of HSV-TK/GCV suicide gene system on cervix cancer cell line Hela in vitro study. Methods: GINaTK retroviral vector containing HSV-TK gene was transduced into PA317 packaging cell by lipofectin. cervix cancer cell line Hela was infected by high titer viral supernatant. Hela /TK cells and Hela cells were used in vitro study. Sensitivity of Hela /TK cells to GCV and bystander effect were observed by MTT test. Results:Hela cells transfered with GINaTK gene displayed a higher tumor cell-killing effect than the parent cells. when Hela /TK cells ratio reached to 10%,the tumor cell-killing proportion was about 50 %. The group of Hela /TK cells exhibiting high sensitivity to GCV, the hallmark of apoptosis as typical mono-and-oligonucleosomal length DNA fragmentation "ladder" could be verified by agrose gel electrophoresis. Conclusion: The result of study shows that the HSV-TK gene can be transducted into cervix cancer Hela cell under the mediation of retrovirus and demonstrates stable expressing, HSV-TK/GCV suicide gene therapy system may improve the antitumoral efficiency . The bystander effect can be observed in HSV-TK/GCV system in in vitro.更多还原