KDR启动子驱动的双自杀基因对实体瘤细胞及血管内皮细胞的靶向杀伤作用

目的腺病毒介导KDR启动子驱动的CDglyTK融合基因(AdKDRCDglyTK)体系对实体肿瘤细胞及血管内皮细胞靶向杀伤作用。方法分别将KDR和CMV启动子融合基因腺病毒体外感染表达KDR的ECV304、MCF7、MGC803及SW620细胞株和不表达KDR的LS174T细胞株,并给予不同浓度的前药5FC和/或GCV,观察其对各细胞株的杀伤效应及其旁观者效应。结果两种腺病毒对各细胞的感染率随腺病毒滴度的递增而增加,当MOI为200时,各细胞株均达约100%感染;感染AdCMVCDglyTK的各组细胞和感染AdKDRCDglyTK的ECV304、MCF7、MGC803及SW620对前药具有较高的敏感性,且其敏感性差异无统计学意义(P均>0.05);与其他细胞相比,感染AdKDRCDglyTK的LS174T细胞对前药不敏感(P<0.01);且观察到该体系明显的旁观者效应。结论KDR基因启动子调控的CDglyTK融合基因体系可选择性杀伤实体肿瘤细胞及血管内皮细胞。

Adenovirus mediated double suicide gene under controlling by KDR promoter kills selectively vein endothelial cells and solid tumor cells

Objective To evaluate the selectively killing effect of adenovirus (Ad) mediated double suicide gene under the regulation of KDR promoter on vein endothelial cells and some sorts of solid tumor cells.Methods KDR producing cells (ECV304,MCF7,MGC803 and SW620) and the KDR nonproducing cells (LS174T) were infected by the two Ads (AdEasy-KDR-CDglyTK and AdEasy-CMV-CDglyTK,),followed by treatment with 5-FC and GCV.Then their killing effects on various kinds of cells were evaluated and their bystander effects were analyzed.Results The infection rate of the two resultant recombinant Ads to all the cells was increased gradually with the addition of multiple of infection (MOI) of Ads.All the cells infected with Ad-CMV-CDglyTK and some cells (ECV304,MCF7,MGC803 and SW620) infected with Ad-KDR-CDglyTK were sensitive highly to the prodrugs,and there were no significant differences among them (P> 0.05).Compared with the others,the LS174T cells infected with Ad-KDR-CDglyTK appeared to be unsensitive to the two prodrugs (P< 0.01).In addition,they all had considerable bystander effect.Conclusion The CD/TK fusion gene driven by KDR promoter has selectively killing effect on the KDR-expressing endothelial cells and solid tumor cells.