国家自然科学基金(30772401)和湖南省卫生厅科研基金(B2007006)资助项目
This work was supported by grants from The National Natural Science Foundation of China (30772401) and Scientific Research Fund of Hunan Provincial Health Department (B2007006)
NPCEDRG基因是采用基因定位候选克隆策略获得的一个鼻咽癌候选抑瘤基因.NPCEDRG在鼻咽癌细胞和组织中表达下调,重新恢复NPCEDRG基因在CNE2细胞系的表达,可部分逆转CNE2 的恶性表型.为揭示NPCEDRG基因在鼻咽癌细胞和组织中表达下调的分子机制,联合应用生物信息学和报告基因载体系统分析方法对NPCEDRG基因启动子区进行克隆及功能分析,系统发育进化足迹分析结果表明,NPCEDRG基因5′端调控区-180~+235 bp区间在脊椎动物中高度保守,该保守区域中存在包括CCAAT/NFY、STAT1和SP1等转录因子结合位点.构建Luc和/或EGFP报告基因表达载体并检测其启动子活性,-146~-8 bp区域有较强的启动子活性,电泳迁移阻滞分析实验(EMSA)提示,CCAAT/NFY转录因子结合位点是NPCEDRG基因的转录调控元件.因此,研究确定-146~-8 bp区域是NPCEDRG基因核心启动子区域且启动子核心元件CCAAT/NFY可能参与NPCEDRG基因的转录调控.
NPCEDRG is an NPC associated suppressive gene cloned by positional candidate cloning strategy. Its transcriptional down-expression has been shown in the cell lines and primary tumor tissues of NPC. Reintroduction of NPCEDRG into CNE2, a cell line derived from NPC, was effective to induce cell differentiation, control cell growth, and regulate the cell cycle. To uncover the molecular mechanisms underlying down-expression of NPCEDRG in NPC cells, bioinformatics approaches and functional assays in different tumor cell lines were used to identify and characterize the NPCEDRG core promoter and cis-acting elements. The conserved region from -180 to +235 bp was found in the potential promoter among 6 vertebrate species by the ECR browser, and there have several potential binding sites for transcription factors, such as CCAAT/NFY, STAT1 and SP1. To characterize the NPCEDRG core promoter, transient luciferase and/or EGFP reporter assay were carried out with the construct pGL3-en138. The results demonstrated that the core promoter is located at the conserved region from -146 to -8 nucleotides. Gel shift assay revealed the specific binding of some nuclear proteins to probes containing a putative CCAAT/NFY site,suggesting that the CCAAT/NFY site contributes to the regulation of NPCEDRG gene expression.
侯德富,关勇军,关 瑞,欧阳咏梅,余艳辉,陈主初.人NPCEDRG基因启动子的克隆及CCAAT/NFY结合位点初步分析[J].生物化学与生物物理进展,2011,38(8):713-723
复制生物化学与生物物理进展 ® 2025 版权所有 ICP:京ICP备05023138号-1 京公网安备 11010502031771号