肺腺癌细胞A549中人survivin基因核心启动子位置与功能初探  

Core Promoter of Human Survivin Gene in Lung Adenocarcinoma Cell Line A549

作　　者：陈斌[1] 陈余清[2] 李伟[2] 

机构地区：[1]上海市肺科医院,上海200433 [2]蚌埠医学院第一附属医院,安徽蚌埠233304

出　　处：《肿瘤学杂志》2008年第11期867-871,共5页Journal of Chinese Oncology

基　　金：国家自然科学基金(30772532);安徽省自然科学基金(070413094)

摘　　要：[目的]探讨人survivin核心启动子的位置及在A549细胞中的转录激活功能。[方法]在生物信息学分析基础上,构建了4个分别由survivin 5′上游-1436^+64、-612^+64、-414^+64和-203^+64片段驱动的报告载体,分别瞬时转染A549及HK-2细胞,观察其在两种细胞的表达活性并比较在A549细胞中表达的活性差异。[结果]生物信息学提示survivin TSS位于ATG上游-64bp处,启动子近端富含若干潜在Sp1、p53和HIF-1α等结合位点。4种报告载体在A549细胞均有活性,且不同长度的启动子活性存在统计学差异。而在HK-2细胞中,4种载体未见显著性差异。[结论]人survivin启动子的活性具有肿瘤特异性,核心启动子可能位于TSS上游-203bp至+64bp处。suvivin核心启动子内可能存在2处HIF-1α的结合位点。[Purpose]To explore the location and transcriptional activity of human survivin gene core promoter in lung adenocarcinoma cell line A549. [Methods] Based on the bioinformatics analysis, four lueiferase reporter vectors were constructed that driven by -1436～＋64, -612～＋64, -414～＋64 and -203～＋64 5＇ upstream uncoding sequences of human survivin gene, transiently transfected into A549 and HK-2 cell line, respectively. The luciferase activities of the four vectors above in A549 or in HK-2 cell line were measured, and discrepances of activites of four vectors in A549 cell line driven by different lengths of uncoding upstream sequences being deleted from 5＇-3＇ were compared. [Results] Bioinformatics assay indicated survivin TSS was located at -64 bp upstream of ATG, the proximal promoter of survivin gene was detected several potential binding sites for Spl, p53 and HIF-1α, etc. The four reporter vectors all presented activity in A549 cell line, and the activity of luciferase driven by different fragments displayed statistical significance. In HK-2 cell, however, there were no significant differences among the four vectors above. [Conclusion] The survivin promoter has tumor-specific activity. The core promoter activity might be located between -203 bp and ＋64 bp. There are 2 putative binding sites of HIF-1α in the core promoter region of human survivin gene.

关 键 词：肺肿瘤 腺癌 A549细胞 SURVIVIN 启动子 

分 类 号：R734.2[医药卫生—肿瘤]