泽漆抑制三阴乳腺癌MDA-MB-231细胞及其凋亡机制研究  

Inhibitory effect of Euphorbia helioscopia on human triple-negative breast cancer MDA-MB-231 cells and its possible mechanism of apoptosis

作　　者：王筱婧[1] 王东兴 范洁[3] 高越[4] 张海[3] 

机构地区：[1]中国药学会国际交流部,北京100022 [2]陆军总医院263临床部,北京101149 [3]同济大学附属第一妇婴保健院,上海201204 [4]第二军医大学药学院实验教学中心,上海200433

出　　处：《药学实践杂志》2017年第4期337-340,358,共5页Journal of Pharmaceutical Practice

摘　　要：目的研究泽漆对三阴乳腺癌MDA-MB-231细胞的作用及其作用机制。方法用MTT法检测细胞活力;用荧光显微镜法测定MDA-MB-231细胞的活性氧(ROS)生成量;用流式细胞仪检测细胞的凋亡率;用TUNEL检测法检测细胞凋亡DNA碎片;用Western blot检测caspase-9、caspase-3、PARP等凋亡相关因子的水平变化。结果 MTT试验显示泽漆提取物对MDA-MB-231细胞具有显著的抑制作用,但作用可被ROS抑制剂NAC及caspase抑制剂Z-VAD-FMK所消除;荧光显微镜检测显示泽漆提取物能显著提高ROS的生成;流式细胞仪检测显示泽漆提取物处理后,PI染色阳性细胞明显增加,但被NAC减弱。Caspase-9、caspase-3在提取物处理后均转为激活形式,PARP被剪切。TUNEL法显示,提取物处理后细胞凋亡碎片明显增多,而提前加入ROS抑制剂NAC和caspase抑制剂Z-VAD-FMK能使泽漆提取物诱导凋亡的DNA碎片明显减少。结论泽漆乙酸乙酯提取物可以有效抑制MDA-MB-231细胞的生长,诱导其凋亡,其作用机制可能与ROS过量生成所致的线粒体损伤途径有关。Objective To investigate the effect of Euphorbia helioscopia on MDA-MB-231 cells and its mechanism .Methods The cell viability was detected by MTT assay . The production of ROS in MDA-MB-231 cells was measured by fluo- rescence microscopy . The apoptotic rate was detected by flow cytometry . Apoptosis DNA fragments were detected by TUNEL assay . Western blot was used to assess the expression of caspase-9, caspase-3 and PARP. Results MTT assay showed that the extract significantly inhibited the viability of MDA-MB-231 cells,which can be diminished by the ROS inhibitor NAC and the caspase inhibitor Z-VAD-FMIC . The marked increase in the production of ROS induced by the extract was observed with fluorescence microscopy . Flow cytometry showed that the PI positive staining cells increased significantly after the treatment of the extract,but was diminished by NAC . Caspase-9 and caspase-3 were activated after the treatment of the extract while the PARP was cleaved . TUNEL showed that a significant increase in apoptotic DNA fragmentation induced by the extract,which can be diminished by NAC and Z-VAD-FMK . Conclusion Ethyl acetate extract inhibited the MDA-MB-231 cells and inducedapoptosis . The mechanism may involve with the mitochondrial damage due to the excessive ROS .

关 键 词：泽漆 MDA-MB-231细胞 三阴乳腺癌 ROS 线粒体途径 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]