CD-TK双自杀基因系统对前列腺癌细胞的杀伤作用  

Toxic effects of CD-TK double suicidal gene system against prostate carcinoma cells

作　　者：朱文辉[1] 谭万龙[1] 黄河[1] 石向华[1] 谢毅[1] 

机构地区：[1]南方医科大学南方医院泌尿外科,广东广州510515

出　　处：《南方医科大学学报》2007年第4期479-481,484,共4页Journal of Southern Medical University

基　　金：广东省自然科学基金(020059)~~

摘　　要：目的探讨CD-TK融合双自杀基因对前列腺细胞的杀伤作用。方法应用缺陷性腺病毒载体将CD-TK融合双自杀基因以及绿色荧光蛋白基因(GFP)转染RM-1细胞,以GFP是否表达作为间接鉴定转染成功的标志,以RT-PCR鉴定作为是否转染的标准。联合前药5-FC和/或GCV作用于转染了CD-TK基因的RM-1细胞,以MTT法检测自杀基因系统对转染后的RM-1细胞的杀伤作用,以未转染的RM-1细胞作对照。结果以腺病毒感染RM-1细胞72h后,在荧光显微镜下可见每个细胞都能发出淡绿色荧光,表明该基因已转染到RM-1细胞中,RT-PCR可检测到转染双基因的RM-1细胞含有CD-TK基因。与对照组相比较,当GCV浓度为125μg/ml时,转染了CD-TK基因的RM-1细胞的存活率为47.27%;与对照组相比较,当5-FC浓度为160μg/ml时,RM-1细胞的存活率为71.56%;而两种前药联合应用时RM-1细胞的存活率为18.45%,低于单一用药组(P<0.05)。结论CD/5-FC、TK/GCV自杀基因系统对前列腺癌细胞具有较强的杀伤作用,而应用CD-TK融合自杀基因系统明显优于单一的自杀基因系统。Objective To evaluate the toxic effects of the CD-TK fusion gene systems against prostate carcinoma cell line RM-1 for assessing the value of suicidal gene therapy for prostate carcinoma. Methods CD-TK fusion gene and green fluorescent protein （GFP） gene were transfected into RM-1 cells through adenovirus vectors. RT-PCR was used to demonstrate successful transfection and transcription of the suicidal genes. The toxic effects of 5-FC and GCV used alone or in combination on the transfected cells were observed by MTT assay, with the non-transfected RM-1 cells serving as control. Results Cytotoxic activity of CD/5-FC and TK/GCV systems against RM-1 cells was observed, and combined treatment with the two drugs resulted in significantly lowered survival of CD-TK-expressing cells （P〈0.05）. After exposure to 5-FC and GCV for 72 h, the survival rate of the transfected cells decreased to 71.56% and 47.27%, respectively, and their combined use resulted in a survival rate as low as 18.46%. Conclusion CD-TK fusion double suicidal gene system can produce significantly stronger toxic effect against RM-1 cells in vitro than either of suicidal genes.

关 键 词：自杀基因 前列腺癌 基因治疗 

分 类 号：R737.25[医药卫生—肿瘤]