Involvement of transcription factor activator protein-2α in doxazosin-induced HeLa cell apoptosis  

作　　者：Lu GAY Dong-xing ZHU Li-ping YANG Ru-shi LIU Feng YAN Jian ZHANG 

机构地区：[1]The Key Laboratory of Protein Chemistry and Developmental Biology, Ministry of Education of China, College of Life Sciences, Hu-nanNormal University, Changsha 410081, China

出　　处：《Acta Pharmacologica Sinica》2008年第4期465-472,共8页中国药理学报（英文版）

基　　金：This work was supported in part by the 973 project of the Ministry of Science and Technique of China （№ 2005CB522505 and 2006CBOF0306）, the National Natural Science Foundation of China （№ 20335020 and 30470945）, the Program for Changjiang Scholars and Innovative Research Team in University （№ IRT0445）, and the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China （№ 705041）.

摘　　要：Aim： To investigate the pro-apoptotic effects of α- 1-adrenergic inhibitor doxazosin in HeLa cells and the potential involvement of transcription factor activator protein-2α （AP-2α） in doxazosin-induced apoptosis. Methods： The HeLa cells were exposed to various concentrations of doxazosin for 16 h. Apoptosis was detected using a DNA fragmentation assay, Hoechst 33258 staining, and flow cytometric analysis. The expression of AP-2α and caspase-3 was detected by relative quantitative RT-PCR and Western blot assays, respectively. After the respective transfections of the HeLa cells with AP-2α overexpressing constructs and an antisense oligonucleotide against AP-2α, apoptosis was assessed by flow cytometric analysis, and the expression of AP-2α and caspase-3 was detected by relative quantitative RT-PCR and Western blot assays. The colorimetric assay was performed to detect the caspase-3 activity. Results： Treatment with various concentrations of doxazosin for 16 h increased the apoptotic rate and total cell death rate of the HeLa cells in a dose-dependent manner and upregulated the expression of AP-2α and caspase-3 in a dose-dependent manner. A dose-dependent increase was observed in the caspase-3 activity. Overexpressing AP-2α led to the increased rate of doxazosin-induced apoptosis and the total cell death, whereas doxazosin-induced apoptosis and the total cell death in HeLa cells decreased by antisense AP-2α. Furthermore, overexpressing AP-2α increased the expression and activity of caspase-3, whereas antisense AP-2α in part abolished the increased effects of doxazosin on caspase-3 expression and activity. Conclusion： Doxazosin induces apoptosis in HeLa cells in a dose-dependent manner, and transcription factor AP-2α is functionally involved in doxazosin-induced HeLa cell apoptosis.Aim： To investigate the pro-apoptotic effects of α- 1-adrenergic inhibitor doxazosin in HeLa cells and the potential involvement of transcription factor activator protein-2α （AP-2α） in doxazosin-induced apoptosis. Methods： The HeLa cells were exposed to various concentrations of doxazosin for 16 h. Apoptosis was detected using a DNA fragmentation assay, Hoechst 33258 staining, and flow cytometric analysis. The expression of AP-2α and caspase-3 was detected by relative quantitative RT-PCR and Western blot assays, respectively. After the respective transfections of the HeLa cells with AP-2α overexpressing constructs and an antisense oligonucleotide against AP-2α, apoptosis was assessed by flow cytometric analysis, and the expression of AP-2α and caspase-3 was detected by relative quantitative RT-PCR and Western blot assays. The colorimetric assay was performed to detect the caspase-3 activity. Results： Treatment with various concentrations of doxazosin for 16 h increased the apoptotic rate and total cell death rate of the HeLa cells in a dose-dependent manner and upregulated the expression of AP-2α and caspase-3 in a dose-dependent manner. A dose-dependent increase was observed in the caspase-3 activity. Overexpressing AP-2α led to the increased rate of doxazosin-induced apoptosis and the total cell death, whereas doxazosin-induced apoptosis and the total cell death in HeLa cells decreased by antisense AP-2α. Furthermore, overexpressing AP-2α increased the expression and activity of caspase-3, whereas antisense AP-2α in part abolished the increased effects of doxazosin on caspase-3 expression and activity. Conclusion： Doxazosin induces apoptosis in HeLa cells in a dose-dependent manner, and transcription factor AP-2α is functionally involved in doxazosin-induced HeLa cell apoptosis.

关 键 词：activator protein-2α or-1-adrenergic inhibitor APOPTOSIS caspases cervical cancer DOXAZOSIN HeLa cells transcription factors 

分 类 号：R737.33[医药卫生—肿瘤]