HeLa细胞中ezrin基因主要转录调控区的定位分析  

Site-directed analysis of key transcriptional regulatory regions of ezrin gene in HeLa cells

作　　者：高书颖[1] 许丽艳[2] 崔磊[1] 周飞[1] 郭张燕[1] 蔡唯佳[2] 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,汕头515041 [2]汕头大学医学院肿瘤病理研究室,汕头515041

出　　处：《肿瘤》2008年第2期101-103,共3页Tumor

基　　金：国家自然科学基金资助项目(编号:30672376;30570849;30370641);国家教育部博士点基金项目(编号:20050560002);广东省自然科学基金重点项目(编号:05104541)

摘　　要：目的:鉴定ezrin基因在HeLa细胞中的主要转录调控区,初步研究ezrin基因的转录调控机制。方法:采用嵌套缺失技术构建一系列以ezrin基因翻译起始位点上游不同长度DNA序列作为报告基因启动子的重组pGLB质粒,利用双荧光素酶报告基因分析系统检测启动子的转录活力,并利用在线分析程序预测ezrin基因主要转录调控区的潜在转录因子结合位点。结果:获得一系列含不同长度ezrin基因5′侧翼区序列的重组pGLB质粒;当ezrin基因5′侧翼区序列从-1324截短到-890时,转录活力降低约80%,从-146截短到-32时,转录活力几乎完全丧失;在-1324/-890和-146/-32区,存在大量的Sp1转录因子结合位点。结论:在HeLa细胞中,ezrin基因的-1324/-890和-146/-32区是主要的转录调控区。Sp1可能是调控基因转录的重要转录因子。Objective：To identify key transcriptional regulatory regions of ezrin gene in HeLa cells, thus this work maked a preliminary study for revealing its transcriptional regulatory mechanism. Methods： The recombinant pGLB plasmids containing different lengths of DNA fragments upstream of translation initiation site of ezrin gene as reporter promoters were constructed using nested-deletion method, and the promoter activities were detected using dual-luciferase reporter assay system. The potential transcription factor binding sites at key transcriptional regulatory region of ezrin gene were predicted using on-line analyzing programme. Results： The recombinant pGLB plasmids containing different lengths of 5＇-flanking region of ezrin gene were obtained. When the lengths of ezrin 5＇-flanking region were reduced from -1 324 to -890, the transcriptional activity decreased by about 80%. If the length of 5＇-flanking region were deleted from -146 to -32, the transcriptional activities were nearly abolished. Sp 1 transcriptional factor binding sites were ubiquitous at the -1 324/-890 and -146/-32 regions. Conclusion： The -1 324/-890 and -146/-32 regions were two key transcriptional regulatory regions of ezrin gene in HeLa cells. Sp 1 may be an important factor for regulating the transcription of ezrin gene.

关 键 词：肿瘤 实验性 调节元件 转录 转录因子 荧光素酶类 萤火虫 基因 ezrin 细胞 HeLa 

分 类 号：R73-354[医药卫生—肿瘤]