Gan Lu, Zhu Dong-xing, Yang Li-ping, Liu Ru-shi, Yan Feng, Zhang Jian
The Key Laboratory of Protein Chemistry and Developmental Biology, Ministry of Education of China, College of Life Sciences, Hu-nan Normal University, Changsha 410081, China.
Acta Pharmacol Sin. 2008 Apr;29(4):465-72. doi: 10.1111/j.1745-7254.2008.00780.x.
To investigate the pro-apoptotic effects of alpha1-adrenergic inhibitor doxazosin in HeLa cells and the potential involvement of transcription factor activator protein-2alpha (AP-2alpha) in doxazosin-induced apoptosis.
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-2alpha 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-2alpha overexpressing constructs and an antisense oligonucleotide against AP-2alpha, apoptosis was assessed by flow cytometric analysis, and the expression of AP-2alpha 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.
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-2alpha and caspase-3 in a dose-dependent manner. A dose-dependent increase was observed in the caspase-3 activity. Overexpressing AP-2alpha 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-2alpha. Furthermore, overexpressing AP-2alpha increased the expression and activity of caspase-3, whereas antisense AP-2alpha in part abolished the increased effects of doxazosin on caspase-3 expression and activity.
Doxazosin induces apoptosis in HeLa cells in a dose-dependent manner, and transcription factor AP-2alpha is functionally involved in doxazosin-induced HeLa cell apoptosis.
研究α1-肾上腺素能抑制剂多沙唑嗪对人宫颈癌HeLa细胞的促凋亡作用以及转录因子激活蛋白-2α（AP-2α）在多沙唑嗪诱导的细胞凋亡中的潜在作用。
将HeLa细胞暴露于不同浓度的多沙唑嗪中16小时。采用DNA片段化分析、Hoechst 33258染色和流式细胞术分析检测细胞凋亡情况。分别通过相对定量逆转录-聚合酶链反应（RT-PCR）和蛋白质免疫印迹法检测AP-2α和半胱天冬酶-3（caspase-3）的表达。在用AP-2α过表达构建体和针对AP-2α的反义寡核苷酸分别转染HeLa细胞后，通过流式细胞术分析评估细胞凋亡情况，并通过相对定量RT-PCR和蛋白质免疫印迹法检测AP-2α和caspase-3的表达。采用比色法检测caspase-3活性。
用不同浓度的多沙唑嗪处理16小时后，HeLa细胞的凋亡率和总细胞死亡率呈剂量依赖性增加，AP-2α和caspase-3的表达也呈剂量依赖性上调。caspase-3活性呈剂量依赖性增加。过表达AP-2α导致多沙唑嗪诱导的细胞凋亡率和总细胞死亡率增加，而反义AP-2α则降低了HeLa细胞中多沙唑嗪诱导的细胞凋亡和总细胞死亡。此外，过表达AP-2α增加了caspase-3的表达和活性，而反义AP-2α部分消除了多沙唑嗪对caspase-3表达和活性的增加作用。
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在功能上参与了多沙唑嗪诱导的HeLa细胞凋亡。
