双氢青蒿素对人胃癌细胞BGC-823细胞增殖、凋亡的影响及机制研究

Effect of dihydroartemisinin on proliferation and apoptosis of human gastric cancer cell line BGC-823 and its mechanisms

目的研究双氢青蒿素(dihydroartemisinin,DHA)对人胃癌细胞BGC-823细胞增殖、凋亡的影响及机制。方法采用噻唑蓝(methylthiazoletetrazolium,MTT)法检测DHA对人胃癌细胞BGC-823增殖的抑制作用;用流式细胞技术、荧光显微镜、透射电子显微镜检测经DHA处理后BGC-823细胞的凋亡率及形态特征的变化;Western印迹方法检测凋亡相关基因Bax、Caspase-3、Caspase-8表达的变化。结果MTT分析表明,DHA作用可明显抑制BGC-823细胞的增殖,抑制率随药物浓度的增加而增大,且随着时间的延长而增大,具有显著的剂量和时间依赖性(P<0.01),半数抑制浓度(IC50值)为3.4μmol/L。流式细胞术检测结果显示,随着DHA药物浓度的增加,凋亡峰愈加明显,并呈现出剂量依赖性(P<0.01)。形态学的观察结果:BGC-823细胞在DHA作用下出现典型的凋亡形态。Western印迹显示:Bax、Caspase-3、Caspase-8蛋白表达随着DHA给药浓度的增加而增高。结论DHA对BGC-823细胞增殖有抑制作用;DHA通过上调Bax、Caspase-3、Caspase-8表达促进BGC-823细胞凋亡。

Objective To investigate the effect of dihydroartemisinin on the growth and apoptosis of human gastric cancer cell line BGC-823.Methods Methylthiazoletetrazolium(MTT)assay was used to determine the inhibitory rate of dihydroartemisinin with different concentrations on the proliferation of BGC-823 cells.The apoptosis rate and morphological changes of BGC-823 cells treated with dihydroartemisinin were detected by flow cytometry(FCM),fluorescence microscope,transmission electron microscope.Western-blot were used to detect the changes of Bax,Caspase-3,Caspase-8 protein levels in BGC-823 cells.Results MTT assay showed that the proliferation of BGC-823 cells was markedly inhibited after treatment with dihydroartemisinin,and the inhibition rate increases with the increase of drug concentration,and with the extension of time.It has a significant dose and time dependence(P<0.01)and IC50 was 3.4μmol/L.The results of flow cytometry showed that with the increase of drug concentration,the apoptotic peak became more obvious and showed a dose-dependent(P<0.01).Typical apoptotic morphology were observed in BGC-823 cells after induced by dihydroartemisinin.Western blot showed that the expressions of Bax,Caspase-3 and Caspase-8 proteins increased with the increase of dihydroartemisinin concentration.Conclusion The result suggested that dihydroartemisinin could inhibit the proliferation of BGC-823 cells.Dihydroartemisinin promotes the apoptosis of BGC-823 cells by up-regulating the expression of Bax,Caspase-3 and Caspase-8.