双自杀基因重组腺病毒对胃癌细胞及血管内皮细胞的体外特异性杀伤作用  

Specific killing effects of adenovirus-mediated double suicide gene driven by KDR promoter on venous endothelial cells and gastric cancer cells in vitro

作　　者：黄宗海[1] 苏国强[1] 俞金龙[1] 厉周[1] 范应方[1] 周广军[1] 宋慧娟[1] 

机构地区：[1]南方医科大学珠江医院,广东广州510282

出　　处：《第一军医大学学报》2005年第8期947-950,共4页Journal of First Military Medical University

基　　金：国家863计划项目(2001AA217171);广东省自然科学基金项目(013072)~~

摘　　要：目的研究腺病毒介导的KDR启动子驱动CD/TK融合基因(AdKDR-CDglyTK)对胃癌细胞及血管内皮细胞的选择性杀伤作用。方法选择表达KDR的MGC803细胞、ECV304细胞和不表达KDR的LS174T细胞,用腺病毒重组体AdEasy-KDR-CDglyTK和AdEasy-CMV-CDglyTK感染之,观察其感染效率并以RT-PCR方法检测转基因细胞CDglyTK的表达,然后给予不同浓度的前药5-FC及GCV处理,观察该体系对不同细胞株的杀伤效应。结果两种重组体对各细胞株的感染率相似,其感染率随腺病毒滴度的增高而递增。RT-PCR检测发现:除感染AdKDR-CDglyTK的LS174T细胞外,感染AdCMV-CDglyTK的所有细胞及感染AdKDR-CDglyTK的MGC803细胞、ECV304细胞均有目的基因CDglyTK的表达。以感染复数为100的两种腺病毒分别感染各细胞株,其表现出对前药不同的敏感性:感染AdCMV-CDglyTK的所有细胞株和感染AdKDR-CDglyTK的MGC803细胞、ECV304细胞对前药具有较高的敏感性,且其敏感性差异无显著性意义(P>0.1);相比之下,感染AdKDR-CDglyTK的LS174T细胞对前药不敏感(P<0.001)。双自杀基因的疗效优于任一单自杀基因的疗效(P<0.001)。将感染腺病毒的细胞与未感染细胞以不同比例混合培养,观察到该体系明显的旁观者效应。结论KDR基因启动子可调控双自杀基因系统选择性杀伤表达KDR胃癌细胞及血管内皮细胞。Objective To study the specific killing effect of adenovirus（Ad）-mediated double suicide gene under regulation by KDR promoter on gastric cancer cells and venous endothelial cells in vitro. Methods KDR-expressing MGCS03 and ECV304 cells and non-KDR-expressing LS174T cells were infected by the two Ads （AdEasy-KDR-CDglyTK and AdEasy-CMVCDglyTK）, and expression of CDglyTK was detected by reverse transcriptional （RT） PCR. After treatment with 5-FC and GCV, the killing effects of the double suicide genes on various cells were evaluated. Results The infection rate of the two resultant recombinant Ads did not differ significantly in the cells. RT-PCR demonstrated the presence of CDglyTK gene product in all the cells infected by Ad-CMV-CDglyTK and all but SL147T cells infected by Ad-KDR-CDglyTK. All the cells infected by Ad-CMV-CDglyTK and ECV304 and MGCS03 cells infected Ad-KDR-CDglyTK were highly sensitive to the prodrugs. In contrast, LS 174T cells infected by Ad-KDR-CDglyTK did not appear sensitive to the two prodrugs （P〈0.001）. In addition, the effect of the double suicide gene was much stronger than that of either of the single suicide gene （P〈0.001）, showing also considerable bystander effect. Conclusions The double suicide gene driven by KDR promoter has specific killing effect on KDR-expressing gastric tumor cells and venous endothelial cells in vitro.

关 键 词：肿瘤 血管内皮 KDR启动子 自杀基因治疗 腺病毒 

分 类 号：R735.2[医药卫生—肿瘤]