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目的 观察双自杀基因的共表达与单自杀基因单独表达对癌细胞的杀伤作用。方法 分别构建了以 CMV为启动子 ,含单纯疱疹病毒胸苷激酶（ HSV-TK）和 /或大肠杆菌胞嘧啶脱氨酶（ Ecoli.CD）单、双自杀基因的真核表达载体。在脂质体介导下将基因导入细胞，经 G418筛选出稳定表达的克隆。用 PCR、半定量 RT-PCR检测各组基因的整合及表达。给予前体药物 5-氟胞嘧啶（ 5-flourocytosine， 5-Fc）和 /或无环鸟苷（ Ganciclovir,GCV）后，用 MTT法测定各转基因组细胞的存活率。 结果 单、双自杀基因均在 GLC-82细胞中稳定表达，双基因转染组对细胞增殖的杀伤及旁杀伤效应高于单基因组。 结论 TK＋ CD/5-Fc＋ GCV的双基因共表达体系较 CD/5-Fc或 TK/GCV单基因体系对细胞具有更强的杀伤作用。
Objective To observe the co-expression of double suicide genes and the killing effect of single suicide gene expression alone on cancer cells. Methods The eukaryotic expression vectors containing CMV as the promoter and herpes simplex virus thymidine kinase (HSV-TK) and/or E. coli cytosine deaminase (Ecoli.CD) single and double suicide genes were constructed. The gene was introduced into the cells by liposome and the clones stably expressed were screened by G418. The integration and expression of genes in each group were detected by PCR and semi-quantitative RT-PCR. After administration of the prodrugs 5-fluorourocytosine (5-Fc) and/or Ganciclovir (GCV), the survival rate of each transgenic cell group was measured by the MTT assay. Results The single and double suicide genes were stably expressed in GLC-82 cells. The double-gene transfection group had higher killing and side killing effects on cell proliferation than single gene groups. Conclusion The double-gene coexpression system of TK+CD/5-Fc+GCV has a stronger killing effect on cells than the CD/5-Fc or TK/GCV single gene system.