ezrin基因在几种癌细胞中的表达及其5′侧翼区序列转录活性的鉴定  

Expression of ezrin Gene in Some Carcinoma Cells and Identification of Its 5′-Flanking Transcriptional Activity

作　　者：高书颖[1] 许丽艳[2] 孟令英[1] 崔磊[1] 周飞[1] 李恩民[1] 

机构地区：[1]汕头大学医学院生物化学与分子生物学教研室,广东515041 [2]汕头大学医学院肿瘤病理研究室

出　　处：《肿瘤防治研究》2008年第1期1-4,共4页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金资助项目(30672376;30570849;30370641);国家教育部博士点项目基金资助项目(20050560002);中国博士后科学基金资且项目(20070410846);广东省自然科学基金重点项目(05104541);广东省自然科学基金博士科研启动项目(7301043)

摘　　要：目的检测ezrin基因在几种癌细胞中的表达及其5′侧翼区序列的转录活力,探讨ezrin基因在癌细胞的表达调控机制。方法采用逆转录聚合酶链反应(RT-PCR)及Western blot法检测ezrin基因在食管癌细胞EC109、EC171、EC8712、SHEEC,胃癌细胞N87、BGC823,肺癌细胞A549、95D,肝癌细胞HepG2,白血病细胞K562和宫颈癌细胞HeLa中的mRNA和蛋白表达水平;采用PCR法构建以ezrin基因翻译起始位点上游-1444/+134序列为启动子的真核细胞表达质粒pGLB-hE(-1444/+134);采用双荧光素酶报告基因分析系统检测-1444/+134序列在EC109、Hela、A549和BGC823细胞中的转录活力。结果ezrin基因在食管癌等几种癌细胞中均有高表达,其5′侧翼区-1444/+134序列具有较强的转录活力。结论ezrin基因5′侧翼区序列的转录活性可能对ezrin基因的几种癌细胞中的高表达起重要作用。Objective To investigate the expression of ezrin gene in some carcinoma cells, to detect the transcriptional activity of its 5＇-flanking region, and to elucidate the transcriptional regulatory mechanism of the ezrin gene in carcinoma cells. Methods Expression of ezrin gene in esophageal carcinoma EC109, EC171, EC8712 and SHEEC, gastric carcinoma N87 and BGC823, lung carcinoma A549 and 95D, hepatocellular carcinoma HepG2, leukemia K562, and cervical carcinoma HeLa cells was detected using reverse transcriptase polymerase chain reaction （RT-PCR） and western blot analysis. Plasmid pGLB-hE（ - 1444/＋ 134）, containing - 1444/＋ 134 of ezrin gene fused to the luciferase gene, was generated by PCR method. Transcriptional activity of the - 1444/＋ 134 region in some carcinoma cells was examined using dual-luciferase reporter assay system. Results All the tested carcinoma cells high-expressed ezrin mR- NA and Ezrin protein. - 1444/＋ 134 region of ezrin gene had strong transcriptional activity compared with SV40 promoter （P〈0. 05） in EC109, Hela, A549 and BGC823 cells. Conclusion The transcriptional activity of the ezrin gene 5＇-flanking region may paly an important role in the high-expression of Ezrin in some carcinoma cells.

关 键 词：EZRIN基因 逆转录聚合酶链反应 WESTERN BLOT检测 双荧光素酶报告基因分析 

分 类 号：Q786[生物学—分子生物学]