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

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3.4 μmol/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。

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

ObjectiveTo investigate the effect of dihydroartemisinin on the growth and apoptosis of human gastric cancer cell line BGC-823.MethodsMethylthiazoletetrazolium（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.ResultsMTT 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 3.4 μmol/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.ConclusionThe 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.