【摘要】 探讨双氢青蒿素诱导人胰腺癌JF-305细胞凋亡作用及活性氧在双氢青蒿素诱导JF-305细胞凋亡中的作用。采用MTT法考察不同浓度双氢青蒿素对人胰腺癌JF-305细胞增殖的影响,流式细胞术检测细胞周期,Hochest 333258荧光染色法观察细胞凋亡形态,Annexin V荧光染色法检测JF-305细胞凋亡的变化,DCFH-DA检测凋亡过程中活性氧(ROS)的变化。Western blot检测细胞内Bax,Bcl-2,Cleaved caspase-3,Cleaved caspase-9和Cyto C蛋白表达的变化。与对照相比,双氢青蒿素作用JF-305细胞48 h,细胞增殖受到明显抑制(P<0.05);细胞被阻滞于G2/M期;细胞出现核浓缩聚集、碎裂的凋亡形态,细胞凋亡比例升高(P<0.05);DCFH-DA检测双氢青蒿素给药组细胞ROS明显升高(P<0.05);Western blot结果显示,双氢青蒿素作用后细胞内Bcl-2蛋白表达下调,Bax蛋白表达上调,Bax/Bcl-2蛋白表达比例升高,Cleaved caspase-3,Cleaved caspase-9和Cyto C蛋白表达升高。双氢青蒿素能诱导JF-305细胞凋亡,其凋亡过程可能与ROS的生成增加相关。更多还原

【Abstract】 To investigate the effect of dihydroartemisinin on apoptosis of human pancreatic cancer cell line JF-305 and the role of reactive oxygen species( ROS) in the apoptosis of JF-305 cells induced by dihydroartemisinin. MTT assays were used to detect effect of different concentrations of dihydroartemisinin on cells proliferation of JF-305 lines. Cell cycle was detected by flow cytometry,and the apoptotic morphology was observed by Hoechst 333258 fluorescence staining. Annexin V fluorescence staining was used to detect the apoptosis changes of JF-305 cells,while DCFH-DA was used to detect the changes of ROS during apoptosis process. Western blot was used to detect the protein expression changes of Bax,Bcl-2,Cleaved caspase-3,Cleaved caspase-9 and Cyto C. As compared with the control group,the JF-305 cells proliferation was inhibited significantly( P < 0. 05) after treatment with different concentrations of dihydroartemisimin for 48 h; cell cycle was blocked in the G2/M phase; apoptotic morphology of nuclear condensation,aggregation,and fragmentation was found,and the apoptosis ratio was increased( P < 0. 05). DCFH-DA detection showed that the cell ROS was increased significantly after dihydroartemisinin treatment( P < 0. 05). Western blot results showed that the expression of Bcl-2 protein was down-regulated; the expression of Bax protein was up-regulated; the ration of Bax/Bcl-2 was increased and the protein expression levels of Cleaved caspase-3,Cleaved caspase-9 and Cyto C were increased after dihydroartemisinin treatment. Therefore,dihydroartemisinin could induce apoptosis of JF-305 cells,and the possible mechanism may be related to the formation and increasing of ROS.更多还原