摘要
目的探讨双氢青蒿素(dihydroartemisinin,DHA)对人前列腺癌细胞系PC-3的凋亡诱导作用,并探讨其可能机制。方法人前列腺癌PC-3细胞经不同浓度(0、25、50和100μmol/L)DHA处理48 h,用FCM法检测各组细胞凋亡率。用荧光定量PCR检测细胞中HSP70 mRNA的表达。用蛋白质印迹法检测细胞中HSP70蛋白、凋亡酶激活因子(Apaf-1)及caspase-3的表达;荧光定量PCR及蛋白质印迹法增加两组,即100μmol/L HSP70抑制剂槲皮素(quercetin)作为阳性药物对照组,以DMSO作为溶剂对照组。结果 DHA能明显诱导PC-3细胞凋亡(P<0.05)。不同浓度DHA能明显下调HSP70 mRNA及蛋白表达水平(P<0.05),上调Apaf-1及caspase-3蛋白表达水平(P<0.05)。结论双氢青蒿素能诱导前列腺癌PC-3细胞凋亡,其作用机制可能是DHA干扰HSP70的表达,促进caspase信号通路中Apaf-1及caspase-3表达。
Objective To investigate the induction of dihydroartemisinin（DHA） on prostate cancer cell line PC-3 apoptosis and potential mechanisms. Methods PC-3 cells were incubated with different concentrations （0, 25, 50, and 100 μ mol/L） of DHA for 48 h. The apoptosis was examined by flow cytometry. HSP70 mRNA expression level were determined by RT-qPCR. HSP70, Apaf-1 and caspase-3 protein expressions were determined by Western blot. Two more groups were tested with RT-qPCR and Western blot, 100 μmol/L HSPT0 inhibitor group and the DMSO group. Results DHA can significantly induce the apoptosis of PC- 3 cells （ P 〈 0. 05 ）. The expressions of HSP70 mRNA and protein significantly decreased （P 〈 0. 05 ） , while the expressions of Apaf-1 and caspase-3 pro- teins increased （P 〈 0. 05 ）. Conclusions DHA significantly inhibits the apoptosis of PC- 3 cells in vitro. The mechanisms may be explained by reduced expression HSP70 and the induced expression of Apaf-1 and caspase-3 by DHA.
出处
《基础医学与临床》
CSCD
2016年第9期1232-1236,共5页
Basic and Clinical Medicine
基金
重庆市科技攻关计划(cstc2012gg-yyjs10017)
