NFY结合位点缺失下调NPCEDRG基因启动子转录活性和效率  

Deletion of NFY Binding Site Decreases the Transcriptional Activity and Efficiency of NPCEDRG Promoter

作　　者：侯德富[1] 关瑞[2] 万恂恂[1] 陈主初[2] 

机构地区：[1]湖南师范大学医学院生物化学与分子生物学教研室,长沙410013 [2]中南大学湘雅医院卫生部肿瘤蛋白质组学重点实验室,长沙410008

出　　处：《中国生物化学与分子生物学报》2013年第12期1159-1165,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：湖南省自然科学基金项目(No.13JJ3059);湖南师范大学博士启动基金(No.130618)~~

摘　　要：NPCEDRG基因是一个鼻咽癌(nasopharyngeal carcinoma,NPC)相关基因.NPCEDRG基因启动子为TATA-less启动子,其核心启动子区域位于-146～-8 bp区,该区域包含NFY、STAT1和cMYB等转录因子结合位点.前期研究结果提示,转录因子NFY可能参与NPCEDRG基因的转录调控.为明确NFY转录因子在NPCEDRG基因转录中的作用,本研究应用报告基因载体系统分析方法对NPCEDRG基因核心启动子区进行NFY结合位点(或CCAAT-box)缺失突变及其功能分析,分别构建NPCEDRG基因核心启动子区NFY结合位点缺失突变的Luc和EGFP报告基因表达载体.比较核心启动子和NFY结合位点缺失突变体报告基因载体的转录活性和效率.结果表明,在MCF7和CNE2细胞中,NFY结合位点缺失突变体的转录活性分别约为其核心启动子转录活性66.94%和75.72%,即NFY结合位点缺失致使该启动子转录效率在MCF7和CNE2细胞中分别降低了33.06%和24.28%;EGFP报告基因表达载体系统研究结果与Luc报告基因载体系统结果基本吻合.本研究再次证实,转录因子NFY通过结合NPCEDRG基因启动子的核心元件NFY结合位点参与NPCEDRG基因的转录调控,并提高其基因转录活性和效率.NPCEDRG is a gene discovered to associate with nasopharyngeal carcinoma （NPC）. The down-expression of NPCEDRG transcription has been reported in NPC cell lines and primary tumor tissues. The NPCEDRG promoter is TATA-less, and core sequence is located at -146 to -8 with several potential transcription factor binding sites, including NFY, STAT1 and cMYB. To characterize the role of NFY in NPCEDRG expression, we constructed luciferase or EGFP reporters without the NFY binding site/CCAAT-box （ - 136 - - 120 bp） of the core promoter and used for cell transfection. The deletion of NFY binding site reduced the luciferase activity by 33.06% and 24.28% in MCF7 and CNE2 celllines, respectively. Similar results were found using EGFP reporters. It suggested that the NFY was necessary for normal transcriptional activity of NPCEDRG.

关 键 词：NPCEDRG基因 核心启动子 转录活性 NFY结合位点 

分 类 号：Q71[生物学—分子生物学] R73[医药卫生—肿瘤]