Survivin启动子介导的胞嘧啶脱氨酶(CD)自杀基因的抗肿瘤研究  

Anti-tumor effect of cytosine deaminase suicide gene driven by the Survivin promoter

作　　者：邵红伟[1] 陈辉[1] 李家明[1] 沈晗[1] 吴凤麟[1] 王辉[1] 黄树林[1] 

机构地区：[1]广东药学院生命科学与生物制药学院/广东省生物技术候选药物研究重点实验室,广东广州510006

出　　处：《广东药学院学报》2015年第3期378-382,392,共6页Academic Journal of Guangdong College of Pharmacy

基　　金：国家自然科学基金项目(31100664;31300737;81303292);广东省自然科学基金项目(10151022401000024;2014A030313586)

摘　　要：目的探讨Survivin启动子介导的CD/5-FC自杀系统的抗肿瘤效果。方法利用Survivin启动子替换p EGFP-N1载体上的巨细胞病毒(CMV)启动子,构建重组表达载体p Sur P-EGFP,通过报告基因绿色荧光蛋白(GFP)的表达确定Survivin启动子的特异性;将酿酒酵母的胞嘧啶脱氨酶(CD)基因克隆在p Sur P-EGFP中,构建成重组载体p Sur P-CD,转染肿瘤细胞后利用毛细管电泳仪检测前药5-氟胞嘧啶(5-FC)的转化效率,同时分析肿瘤细胞的凋亡。结果 GFP的表达表明Survivin启动子具有较好的肿瘤靶向性;毛细管电泳检测表明CD能够有效地将5-FC转化为5-FU,显微镜观察和流式分析表明了Survivin启动子介导的CD的抗肿瘤效果。结论 Survivin启动子能有效地介导CD自杀基因的表达,具有较好的靶向性抗肿瘤效果。Objective To investigate the anti-tumor effect of cytosine deaminase（ CD） /5-Flucytosine（ 5-FC） suicide system driven by the Survivin promoter. Methods The cytomegalovirus（ CMV） promoter in p EGFP-N1 was replaced by the Survivin promoter to construct recombinant plasmid p Sur P-EGFP. The expression of the Survivin promoter was determined by the report gene of green fluorescent protein（ GFP）.The CD gene was amplified from Saccharomyces cerevisiae and inserted into p Sur P-EGFP to construct recombinant plasmid p Sur P-CD. The CD gene driven by the Survivin promoter was transferred into tumor cells,and the conversion efficiency of 5-FC by CD was detected with capillary electrophoresis. The apoptosis of tumor cells was also analyzed. Results GFP expression implied the tumor-targeting effect of the Survivin promoter. The capillary electrophoresis analysis showed that 5-FC could be converted by CD. The more obvious apoptosis in Hep G2 cells than that in HEK-293 cells indicated the anti-tumor effect of CD driven by the Survivin promoter. Conclusion The Survivin promoter could mediate the expression of CD in tumor cells and display the anti-tumor targeting effect.

关 键 词：SURVIVIN启动子 自杀基因 胞嘧啶脱氨酶 5-氟胞嘧啶 

分 类 号：R730.5[医药卫生—肿瘤]