Open AccessInvolvement of transcription factor activator protein‐2α in doxazosin‐induced HeLa cell apoptosis 1
Author(s) -
GAN Lu,
ZHU Dongxing,
YANG Liping,
LIU Rushi,
YAN Feng,
ZHANG Jian
Publication year - 2008
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2008.00780.x
Subject(s) - doxazosin , apoptosis , hela , microbiology and biotechnology , dna fragmentation , programmed cell death , biology , activator (genetics) , western blot , cell cycle , cell , chemistry , biochemistry , endocrinology , gene , blood pressure
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.