转录因子AP-2α对HeLa细胞内14-3-3σ表达的影响  

Effect of transcription factor AP-2α on expression of 14-3-3σ in HeLa cells

作　　者：甘露[1] 黄明元[2] 蒙子君[1] 张哲[1] 罗炳德[1] 

机构地区：[1]南方医科大学公共卫生与热带医学学院预防医学实验教学中心,广州510515 [2]广东医学院公共卫生学院劳动卫生与环境卫生教研室,广东东莞523808

出　　处：《重庆医学》2012年第17期1680-1683,共4页Chongqing medicine

基　　金：国家自然科学基金资助项目(31070687);广东省自然科学基金资助项目(9451051501002535)

摘　　要：目的探讨转录因子AP-2α对HeLa细胞内14-3-3σ表达的影响。方法在HeLa细胞内过表达AP-2α或干扰AP-2α的表达,用荧光定量RT-PCR和Western blot检测Hela细胞AP-2α和14-3-3σ的表达量;采用计算机软件预测14-3-3σ的启动子区域的转录因子结合位点;采用双荧光素酶报告基因分析试验验证AP-2α对14-3-3σ启动子转录活性的影响。结果在HeLa细胞内过表达AP-2α后,14-3-3σ的mRNA和蛋白表达水平增高,而用siRNA干扰AP-2α后,14-3-3σ的mRNA和蛋白表达水平降低;软件分析结果显示在14-3-3σ启动子区域存在多个潜在的AP-2α结合位点;双荧光素酶报告基因分析试验证明在HeLa细胞内,AP-2α增强14-3-3σ启动子的转录活性。结论 AP-2α正调控HeLa细胞内14-3-3σ的表达,这种调控作用可能是通过AP-2α增强14-3-3σ启动子的转录活性来完成的。Objective To investigate the effect of AP-2a on thel4-3-3σ expression in HeLa cells. Methods AP-2σ was overexpressed or RNA interfered in HeLa cells. The quantitative RT-PCR and Western blot were performed to examine the expression of AP-2σ and 14-3-3σ. The transcription factor binding sites in the 14-3-3σ promoter were predicted by using the computer software. Dualuciferase reporter assay was used to test the effect of AP-2σ on the transcriptional activity of 14-3-3σ promoter. Results Overexpression of AP-2σ led to the increased mRNA and protein levels of 14-3-3σ, and the mRNA and protein expression of 14-3-3σ was decreased when AP-2σ was interfered by siRNA in HeLa cells. The results from the computer software analysis showed the multiple potential AP-2σ binding sites in 14-3-3σ promoter. Dual-luciferase reporter assay confirmed that AP-2σ enhanced the transcriptional activity of 14-3-3σ promoter. Conclusion AP-2σ positively regulates the 14-3-3σ expression in HeLa cells probably through enhancing the transcriptional activity of 14-3-3σ promoter.

关 键 词：AP-2 14-3-3σ HELA细胞 转录因子 

分 类 号：R363[医药卫生—病理学]