慢病毒介导的双自杀基因诱导胃癌细胞凋亡的研究  

Effect of lentivirus containing double suicide gene on apoptosis of human gastric carciuoma cells

作　　者：孔恒[1] 黄宗海[2] 陈海金[2] 陶霖玉[1] 齐柯[1] 

机构地区：[1]深圳市南山区人民医院甲乳外科,518052 [2]南方医科大学附属珠江医院普通外科

出　　处：《中华实验外科杂志》2010年第1期53-55,共3页Chinese Journal of Experimental Surgery

基　　金：国家重点基础研究863计划资助项目（2001AA217171）

摘　　要：目的 观察慢病毒介导的KDR启动子驱动的CD/TK双自杀基因体系对胃癌细胞SGC-7901的体外杀伤作用.方法 用重组慢病毒FGW-KDRP-CD/TK体外感染SGC-7901细胞.噻唑蓝（MTT）比色法检测前药对细胞的杀伤效应;透射电镜及流式细胞术观察细胞凋亡及细胞内DNA含量.结果 慢病毒的感染率随病毒滴度的增高而递增.当前药更昔洛韦（GCV）为100 mg/L、5-氟胞嘧啶（5-FC）为320 mg/L时,对未转基因及转基因细胞的抑制率分别为18%、94%,两者筹异有统计学意义（P〈0.01）.电镜下用药组出现细胞凋亡.流式细胞仪检测显示,随着药物浓度的增加,凋亡细胞所占的比例逐渐增加.结论 KDR启动子可调控双自杀基因系统选择性杀伤SGC-7901细胞,并且该体系可以诱导胃癌细胞凋亡.Objective To evaluate the killing effects of lentivirus-mediated CD/TK fusion gene controlled by KDR promoter on human gastric carcinoma cells SGC-7901 in vitro.Methods The SGC-7901 cells were infected by the FGW-KDRP-CD/TK.The killing effects of the predrug on SGC-7901 cells were evaluated by MTT method.The apoptosis and cellular DNA content were assayed by electron microscopy and flow cytometry（FCM）.Results The transfection efficiency in SGC-7901 cells Was increased with the increasing adenoviral titer.When the concentrations of the prodrug Ganciclovir（GCV）and 5-FC were 100 mg/L and 320 mg/L,inhibition rate of non-transgenic cells and the genetically modified cells reached 18%and 94%,respectively.There was significant difference between two groups（P〈0.01）.The features of apoptotic cells appeared in the treatment group under electron microscopy.Flow cytometry （FCM） showed the percentage of apeptotie cells Was increased with the increasing drug concentration.Conclusion KDR promoter may regulate the CD/TK fusion gene system to selectively kill the SGC-7901 cells and induce the apoptosis.

关 键 词：胃癌 自杀基因 慢病毒 脱噬作用 

分 类 号：R735[医药卫生—肿瘤]