摘要
目的研究α1-肾上腺素受体拮抗剂多沙唑嗪对HeLa细胞凋亡的影响以及转录因子激活蛋白-2α(activator protein-2α,AP-2α)在多沙唑嗪诱导HeLa细胞凋亡中发挥的作用。方法HeLa细胞经过多沙唑嗪处理、过表达AP-2α或反义寡核苷酸干扰AP-2α表达后,通过流式分析检测凋亡;以相对定量PCR和Western blot分析检测AP-2α和caspase-3的表达。结果多沙唑嗪以剂量依赖性方式增加HeLa细胞的凋亡率和总细胞死亡率,上调AP-2α和caspase-3的表达。AP-2α的过表达导致多沙唑嗪诱导的HeLa细胞的凋亡率和总细胞死亡率增加,增强caspase-3的表达;而反义AP-2α却导致多沙唑嗪诱导的HeLa细胞的凋亡率和总细胞死亡率降低,部分地阻断了多沙唑嗪对caspase-3表达的增强效应。结论多沙唑嗪以剂量依赖性方式诱导HeLa细胞凋亡,转录因子AP-2α在多沙唑嗪诱导的HeLa细胞凋亡中发挥了作用。
Objective To investigate the HeLa cells and the role of transcription factor pro-apoptotic effects of an α1-adrenergic inhibitor, doxazosin, in activator protein-2α （AP-2α） in the process. Methods The HeLa cells were treated with doxazosin （0, 10, 40 or 60 μmol/L） for 16 h, and cell apoptosis was detected by flow cytometric analysis. After doxazosin （60 μmol/L） treated the HaLa cells, the cells transfected with AP-2α overexpressing constructs or an antisense oligonucleotide against AP-2α for 16 h, the apoptosis was analyzed with FCM. The expression of AP-2α and caspase-3 in above 3 groups of cells was detected by relative quantitative RT-PCR and Western blot analysis, respectively. Results Doxazosin increased the apoptotic rate and to- tal cell death rate of the HeLa cells, and upregulated the expressions of AP-2α and caspase-3 in a dose-dependent manner. Overexpressing AP-2α improved the increased apoptotic rate and total cell death rate induced by doxazosin, and enhanced the increased expression of caspase-3. Whereas doxazosin-induced apoptosis and the total cell death in HeLa ceils were decreased by antisense AP-2α, and antisense AP-2α in part abolished the increased effects of doxazosin on caspase-3 expression. Conclusion Doxazosin induces apoptosis in HeLa cells in a dose-dependent manner, and transcription factor AP-2α is functionally involved in this process.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2009年第24期2414-2417,共4页
Journal of Third Military Medical University
基金
南方医科大学公共卫生与热带医学学院院长基金(GW200806)~~
