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［目的］观察逆转录病毒载体／单纯疱疹病毒胸腺嘧啶核苷激酶基因／羟甲基无环鸟苷（ＲＶ／ＨＳＶ１－ｔｋ／ＧＣＶ）基因治疗系统对卵巢癌细胞的体外作用及其“旁观者效应”，并探索提高携有ＨＳＶ１－ｔｋ基因的假型逆转录病毒对卵巢癌细胞的感染率的有效途径。［方法］①以ＶＰＣ／ＨＳＶ１－ｔｋｃ细胞培养液的滤过液重复多次感染Ａ０细胞，然后一组以Ｇ４１８筛选阳性克隆，另一组以ＧＣＶ作杀伤试验。②按不同比例分别将ＡＯ／ＨＳＶ１－ｔｋｃ及ＶＰＣ／ＨＳＶ１－ｔｋｃ与ＡＯ细胞混合培养后加入ＧＣＶ， ５天后消化计数。［结果］①随着ＲＶ感染次数增多，所获得的Ｇ４１８筛选阳性克隆数也明显增多，同时，ＧＣＶ对ＡＯ细胞的杀伤效应也有所提高。②在 ＡＯ细胞群体中如 ＨＳＶ１－ｔｋｃ阳性细胞占 ８０％，则 ＧＣＶ对细胞群体的抑制率可达９５．２％。③当 ＡＯ与 ＶＰＣ／ＨＳＶ１－ｔｋｃ以１：４混合培养时， ＧＣＶ对细胞的生长抑制率为９６．６２％。［结论］ ＧＣＶ杀伤ＡＯ／ＨＳＶ１－ｔｋｃ细胞时对ＡＯ细胞具有一定的“旁观者效应”；重复ＲＶ感染及与ＶＰＣ／ＨＳＶ１－ｔｈｃ细胞混合培养可以明显提高ＲＶ／ＨＳＶ１－ｔｋｃ／ＧＣＶ系统对卵巢癌细胞的作用效果。
[Objective] To observe the effect of RV / HSV1-TK / GCV gene therapy system on ovarian cancer cells in vitro and its “bystander Effect ”and explore an effective way to increase the infection rate of ovarian cancer cells by pseudotyped retrovirus carrying HSV1-tk gene. [Method] ① A0 cells were infected repeatedly with the filtrate of VPC / HSV1-tkc cell culture solution, then one group screened positive clones by G418 and the other group was killed by GCV. ② The AO / HSV1-tkc and VPC / HSV1-tkc were mixed with AO cells by different proportions and then added to GCV. The digestion counts were counted after 5 days. [Results] ①With the increase of the number of RV infections, the number of positive clones screened by G418 also increased significantly. At the same time, the killing effect of GCV on AO cells also increased. ② In the AO cell population such as HSV1-tkc positive cells accounted for 80% of the cell population GCV inhibition rate of up to 95.2%. ③ When AO and VPC / HSV1-tkc were mixed in a ratio of 1: 4, the inhibition rate of GCV to the cells was 96.62%. [Conclusion] GCV killed AO / HSV1-tkc cells have certain “bystander effect” on AO cells; repeated RV infection and mixed culture with VPC / HSV1-thc cells can significantly improve the ovarian Effect of cancer cells.