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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 gene is a NPC related gene.NPCEDRG gene promoter is TATA-less promoter and its core promoter region is located in -146 ~ -8 bp region, which contains NFY, STAT1 and cMYB, etc. Transcription factor binding sites.Previous studies suggest that the transcription factor NFY may be involved in the transcriptional regulation of NPCEDRG gene.To clarify the role of NFY transcription factor in NPCEDRG gene transcription, this study uses reporter gene vector system analysis method of NPCEDRG gene core promoter (Or CCAAT-box) deletion mutation and its function analysis to construct the expression vectors of Luc and EGFP reporter gene with deletion mutation of NFY binding site in NPCEDRG gene core region.Compared the expression of core promoter and NFY binding site The results showed that the transcriptional activity of NFY binding site-deleted mutants in MCF7 and CNE2 cells were about 66.94% and 75.72% of their core promoter transcriptional activity respectively, that is NFY The transcriptional efficiency of this promoter was reduced by 33.06% and 24.28% in MCF7 and CNE2 cells, respectively. The EGFP reporter gene vector The results of this study are basically consistent with the results of Luc reporter gene vector system.The study again confirmed that transcription factor NFY participates in the transcriptional regulation of NPCEDRG gene and enhances its gene transcriptional activity and efficiency by binding NFY binding site, which is the core element of NPCEDRG gene promoter.