二氢青蒿素通过线粒体依赖性途径诱导人胃癌细胞凋亡的机制  

Dihydroartemisinin induces apoptosis of human gastric Cancer cells through mitochondrial pathway

作　　者：王爱军 冯俊伟 王红钰 郑宝军 施华 吴肖 

机构地区：[1]河北唐山市工人医院肿瘤外科,063000

出　　处：《中华实验外科杂志》2012年第7期1312-1314,共3页Chinese Journal of Experimental Surgery

摘　　要：目的探讨二氢青蒿素（DHA）体外诱导人胃癌SGC7901细胞凋亡的分子作用机制。方法体外培养人胃癌SGC7901细胞，不同浓度（6．25、12．50、25．00、50．00、100．00μmol／1）DHA作用24、48、72h后，噻唑蓝（MTT）比色法检测细胞增殖；DHA作用人胃癌SGC7901细胞24h后，流式细胞仪检测细胞凋亡率；分光光度法检测细胞半胱氨酰天冬氨酸特异性蛋白酶3（Caspase-3）和Caspase-9活性变化；线粒体膜电位检测试剂盒（JC-1）荧光染色检测线粒体膜电位（△ψm）的改变；Westernblot法分别测定B-细胞淋巴瘤／白血病-2（bel-2）和bax蛋白的表达水平；Westernb]ot检测胞质中细胞色素C（CytoC）的含量变化。结果DHA对SGC7901细胞增殖抑制具有明显量效和时效关系；与对照组（1．88±0．25）比较，不同浓度DHA作用人胃癌SGC7901细胞24h后，细胞凋亡率[（4．21±0．83）％、（6．92±1．26）％、（9．78±2．55）％、（13．93±2．94）％、（16．02±3．83）％]明显增加（P〈0．05）；与对照组（0．091±0.007、0．079±0．006、100．000±0．000）比较，Caspase-3（0．094±0．008、0．196±0．014、0．288±0．017、0．326±0．024、0．491±0．042）和Caspase-9（0．082±0．008、0．154±0．010、0．176±0．012、0．217±0．015、0．268±0．024）活性逐步升高（P〈0．05）；△ψm（74．18±7．84、70．08±6．53、58．66±2．38、48．79±1．31、31．24±0．73）逐渐下降（P〈0．05）；不同浓度DHA处理组bax蛋白表达逐渐升高（P〈0．05），bcl-2表达逐步下降（P〈0．05），bel-2／bax比值（2．37±0．24、1．51±0．21、0．82±0．16、0．64±0．14、0．37±0．07）显著降低，与对照组（3．11±0．28）比较有剂量依赖趋势（P〈0．05）；与对照组（0．18±0．04）比较，不同浓度DHA均能剂量依赖地诱导胞质CytoC表达（0．21±0．05、0．25±0．06、0．38±0．08、0．39±0．09）增加Objective To investigate the molecular mechanism of induction of apoptosis by dihydroartemisinin （DHA） in human gastric cancer SGC7901 cells in vitro. Methods Cultured SGC7901 ceils were treated with DHA at different concentrations （6.25, 12. 5, 25. 0, 50.0, and 100.0 μmol/L） for different durations （24, 48, and 72 h）, and then the proliferation of cells was assayed by using MTF. SGC7901 ceils were treated with DHA at different concentrations for 24 h, and the cell apoptosis rate was analyzed by using flow cytometry （FCM）. The activities of Caspase-3 and Caspase-9 were assessed by col- orimetric assay. The change of mitochondrial trans-membrane potential （AOm） was measured by JC-1 staining. The expression levels of bcl-2 and bax were detected by using Western blotting. Western blotting was also used to reveal the amount of Cyto C in cytosol. Results Various concentrations of DHA inhibited the growth of SGC7901 cells in a dose- and time-dependent manner. After SGC7901 cells were treated with DHA for 24 h, as compared with control group （ 1.88 ±0. 25）, the cell apoptosis rate [ （4. 21 ±0. 83）%, （6. 92 ± 1.26）% , （9. 78 ±2. 55）% , （ 13.93 ±_2. 94）% , （ 16. 02 ±3.83）% ] were increased significantly and the activities of Caspase-3 （0. 094 ± 0. 008, 0. 196 ± 0. 014, 0. 288 ± 0. 017, 0. 326 ± 0. 024, 0. 491 ± 0.042） and Caspase-9 （0.082 ±0.008, 0. 154 ±0.010, 0. 176 ±0.012, 0.217 ±0.015, 0.268 ±0.024） were increased dose-dependently （ P 〈 0. 05 ）. As compared with control group （ 100 ）, △ψm （ 74. 18 ± 7. 84, 70. 08 ± 6. 53, 58.66 ± 2. 38, 48.79 ± 1.31, 31.24 ± 0. 73 ） was declined significantly （ P 〈 0. 05 ）. Western blotting showed that DHA increased bax expression and decreased bcl-2 expression （ P 〈 0. 05 ）. The ratio of bcl-2/bax （2.37 ±0.24, 1.51 ±0.21, 0.82 ±0.16, 0.64 ±0.14, 0.37 ±0.07） was decreased as compared with control group （3.11 ± 0. 28 ）. Western blotting also revealed that the amount of C

关 键 词：二氢青蒿素 胃癌 脱噬作用 线粒体 

分 类 号：R285[医药卫生—中药学] R735.2[医药卫生—中医学]