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-2α was overex-pressed or RNA interfered in HeLa cells. The quantitative RT-PCR and Western blot were performed to examine the expression of AP-2α and 14-3-3σ. The transcription factor binding sites in the 14-3-3σ promoter were predicted by using the computer software. Dual-luciferase reporter assay was used to test the effect of AP-2α on the transcriptional activity of 14-3-3σ promoter. Results Overexpression of AP-2α 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-2α was interfered by siRNA in HeLa cells. The results from the computer software analysis showed the multiple potential AP-2α binding sites in 14-3-3σ promoter. Dual-luciferase reporter assay confirmed that AP-2α enhanced the transcriptional activity of 14-3-3σ promoter. Conclusion AP-2α positively regulates the 14-3-3σ expression in HeLa cells probably through enhancing the transcriptional activity of 14-3-3σ promoter.
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