重组腺病毒介导的KDR-CDglyTK融合基因系统对胃癌细胞增值的影响

目的 探讨腺病毒介导KDR启动子驱动的融合基因体系对人胃癌细胞SCG7901增值的影响.方法 重组腺病毒AdEasy-KDR-CDglyTK体外感染表达KDR的SCG7901细胞株和对照组不表达KDR的HepG2细胞株,并给予不同浓度的前药GCV(ganciclovir)和/或5-FC(5-fluorocytosine),观察该体系对SCG7901细胞生长增值的影响.结果 携带双自杀基因(CDglyTK)和报告基因(GFP)的重组腺病毒载体,感染复数为100时,95%以上的受感染SCG7901和HepG2细胞中有GFP表达.已转染腺病毒的SCG7901和HepG2细胞同未转染的各自细胞在细胞生长方面无显著性差异(P＞0.05).在前药应用下,已转染腺病毒的SCG7901和HepG2细胞表现出对前药不同的敏感性:表达KDR的SCG7901细胞对前药具有较高的敏感性,与前者相比,不表达KDR的HepG2细胞对前药不敏感(P＜0.01).融合基因的疗效优于单一自杀基因(P＜0.01).结论 KDR基因启动子可以调控融合基因体系选择性地杀伤人胃癌SCG7901细胞,抑制其增值.

Effect of KDR recombinant adenovirus containing double suicide gene on proliferation of human stomach adneocarcinoma SCG7901 cells

Objective To study the effect of adenovirus (Ad)-mediated fusion gene system driven by KDR promoter on the proliferation of human stomach adneocarcinoma SCG7901. Methods The KDR-expressing SCG7901 cells and HepG2 cells that did not express KDR were both transfected with AdEasy-KDR-CDglyTK followed by treatment with the prodrugs 5-FC and/or GCV at different concentrations. The killing effects of the transfection on the cells were evaluated. Results The expression of green fluorescent protein (GFP) was observed in 95% of the infected SCG7901 and HepG2 cells with the multiple of infection (MOI) of the Ads of 100. Transfection of SCG7901 and HepG2 cells did not produce significant changes in the cell growth, and the infected cells exhibited different sensitivities to the two prodrug: SCG7901 cells infected with rAd were highly sensitive to the prodrugs, but the infected HepG2 cells were not (P<0.01). The killing effect of CDglyTK fusion gene on the target cells was much stronger than that of either the single suicide gene (P<0.01). Conclusion CDglyTK fusion gene system driven by KDR promoter selectively kills the KDR-CDglyTK SCG7901 cells and inhibits their proliferation.