肿瘤特异性Survivin启动子与甲胎蛋白启动子的活性评价与比较  

Detection and comparison of transcriptonal activities of tumor-specific survivin and alpha-fetoprotein pro- moters in human hepatocellular carcinoma cells

作　　者：况舸[1] 黄爱龙[1] 唐霓[1] 

机构地区：[1]重庆医科大学病毒性肝炎研究所,教育部感染性疾病分子生物学重点实验室,400010

出　　处：《中华肝脏病杂志》2005年第6期440-442,共3页Chinese Journal of Hepatology

基　　金：国家"863"计划(2001AA217121)

摘　　要：目的研究并比较肿瘤特异性Survivin和甲胎蛋白(AFP)基因启动子在不同肝细胞癌细胞系中的转录活性,为肝癌靶向性基因治疗提供依据。方法采用聚合酶链反应法扩增获得Survivin和AFP 基因的启动子片段,并克隆入表达虫荧光素酶基因的报告质粒中,通过测定荧光素酶报告基因的表达情况, 测定Survivin与AFP基因启动子的转录活性,同时采用巨细胞病毒启动子为对照,评价其转录活性水平。结果Survivin启动子在肝癌细胞中具有相当强的活性,其活性水平在高表达AFP的肝癌细胞中为AFP启动子的54～100倍,达到巨细胞病毒启动子活性的16%～21%。结论Survivin启动子在肝癌细胞系中具有较强的启动活性,可望开发成为新的肝癌靶向性基因治疗工具。Objectives To detect and compare the transcriptional activities of tumor-specific survivin and AFP promoters in various hepatocellular carcinoma cell lines and to lay some groundwork for targeting gene therapy in human hepatocellular carcinoma. Methods The fragment of survivin and AFP promoters were acquired by PCR amplification and were cloned into the reporter plasmid pGL3-Basic, which contained a luciferase gene. The constructed eukaryotic expression plasmid pGL3-SUR and pGL3-AFP, in which the expression of the luciferase was derived by survivin or the AFP promoter, were transfected into three HCC cell lines. At 24 hours post transfection (p.t.), the activity of the luciferase was determined with Dual-Luciferase Reporter Assay System. A pGL3-CMV, containing the CMV promoter controlled luciferase gene, was used as a positive control. Results Both survivin and AFP promoters had transcriptional activities in all three HCC cell lines and the transcriptional activity of the survivin promoter was much higher than the AFP promoter (52-98 times) and reached a level of 16%~21% of the transcriptional activity of the CMV promoter. Conclusion Our data reveals that the survivin promoter possesses a high transcriptional activity in all three established HCC cell lines and may serve as a useful tool for transcriptional targeting gene therapy of HCCs.

关 键 词：Survivin 肿瘤特异性 甲胎蛋白启动子 活性评价 荧光素酶报告基因 巨细胞病毒启动子 靶向性基因治疗 聚合酶链反应法 基因启动子 荧光素酶基因 AFP启动子 转录活性 活性水平 AFP基因 启动子活性 肝癌细胞系 表达情况 治疗工具 

分 类 号：R735.7[医药卫生—肿瘤] R730.51[医药卫生—临床医学]