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目的 观察羟甲基无环鸟苷(GCV) 对转导携有Ⅰ型单纯疱疹病毒胸苷激酶基因( HSV1 - tk) 的AO、3AO 细胞外杀伤作用。方法 分别在携有HSV1 - tk 基因的AO、3AO 细胞培养液中加入不同浓度的GCV,绘制剂量效应曲线,并以荧光细胞激活分选仪(FACS) 测定凋亡细胞比例及细胞周期比例,以光镜及电镜观察细胞形态变化,以DNA 荧光染色观察细胞核内染色质变化。结果 GCV 在体外对于携有HSV1 - tk 基因的AO、3AO 细胞的半数抑制浓度均介于10 - 1 ～10 - 2μg/ml 之间,用药后S 期细胞比例显著增高。结论 GCV 在体外可高效杀伤携有HSV1 - tk 基因的卵巢癌细胞。
Objective To observe the cytotoxic effects of hydroxymethyl acyclovir (GCV) on AO and 3AO transduced with herpes simplex virus thymidine kinase gene (HSV1-tk). Methods Different concentrations of GCV were added to AO and 3AO cell culture fluids carrying HSV1-tk gene, and the dose-response curves were plotted. The proportion of apoptotic cells and the ratio of cell cycle were measured by fluorescence cell activation sorter (FACS). The changes of cell morphology were observed by light and electron microscopy. The changes of chromatin in the nucleus were observed by DNA fluorescence staining. Results The median inhibitory concentration of GCV in AO and 3AO cells carrying HSV1-tk gene was between 10 -1 and 10 -2 μg/ml in vitro, and the proportion of cells in S phase was significantly increased after administration. Conclusion GCV can effectively kill ovarian cancer cells carrying HSV1-tk gene in vitro.