人肝癌特异性HSV-TK/GCV重组腺病毒相关病毒质粒的构建与研究  

Construction and Study of Specific HSV-TK/GCV Reconstructed Adeno Associated Virus Plasmid of Human Hepatocellular Carcinoma

作　　者：李志辉[1] 严律南[1] 赵永恒[1] 刘智敏[1] 刘自明[1] 李玉[1] 

机构地区：[1]四川大学华西医院普外科,成都610041

出　　处：《中国普外基础与临床杂志》2004年第3期234-237,共4页Chinese Journal of Bases and Clinics In General Surgery

摘　　要：目的　构建肝癌特异性HSV TK/GCV重组腺病毒相关病毒质粒并了解其在细胞内的表达。方法　以腺病毒相关病毒的质粒WAV2作为载体 ,将TK基因插在甲胎蛋白 (AFP)增强子 /白蛋白启动子 (AFP增强子/ALB启动子 )的调控基因的下游 ,重组成pWAV2 /AFP ALB/HYTK质粒载体。同时构建质粒载体pEGFP 1/AFP ALB。然后将上述两种不同载体分别转入AFP阳性表达的HepG2细胞株以及阴性表达的 772 1、SPC和 790 1细胞株。结果　绿色荧光蛋白只在AFP阳性表达的HepG2细胞株表达 ,采用PCR技术 ,以HSV TK的引物扩增所抽提的总DNA中 ,只有AFP阳性表达的HepG2细胞株扩增出了 710bp的DNA片段。结论　pWAV2 /AFP ALB/HYTK质粒载体在体外实验中具有很好的靶向性。Objective To investigate target gene therapy for hepatocellular carcinoma (HCC). Methods Herpes simplex virus thymidine kinase (HSV TK) gene was inserted into the gene of AFP enhancer/ALB promoter with adeno associated virus (AAV) plasmid (WAV2) as a carrier, and a hybrid plasmid pWAV2/AFP ALB/HYTK was constructed. Besides, plasmid pEGFP 1/AFP ALB was also constructed. Two kinds of plasmids were transferred into AFP positive cells HepG2 and AFP negative cells 7721, SPC and 7901.Results It was found that enhance green fluorescence protein could only be seen in AFP positive cells HepG2. 710 bp DNA was amplified only in AFP positive HepG2 cells.Conclusion Plasmid pWAV2/AFP ALB/HYTK for HCC demonstrates specificity in vitro .

关 键 词：肝癌 特异性HSV-TK/GCV 腺病毒相关病毒 重组质粒 基因治疗 

分 类 号：R735.7[医药卫生—肿瘤]