摘要
目的:观察华蟾素在诱导人乳腺癌MCF-7细胞株凋亡的过程中Bax和Bcl-2蛋白表达的变化及意义。方法:在人乳腺癌细胞株MCF-7中分别加入不同终浓度的华蟾素,24h、48h及72h后检测指标。应用MTT比色法检测细胞增殖活力,倒置显微镜观察细胞形态学变化,TUNEL法检测细胞凋亡指数,Western blot法检测Bax/Bcl-2蛋白表达,共聚焦显微镜观察Bax蛋白分布。结果:华蟾素能显著抑制MCF-7细胞增殖;倒置显微镜下发现凋亡的MCF-7细胞固缩、核染色质凝聚;western blot发现华蟾素能上调MCF-7细胞Bax蛋白表达,但对Bcl-2蛋白表达无影响,正常MCF-7细胞内的Bax均匀分布于细胞质中,华蟾素处理后,Bax明显地聚集于线粒体,呈现线粒体的转位。结论:华蟾素诱导MCF-7细胞凋亡,并在该过程中导致的Bax蛋白表达上调并且向线粒体转位。
Objective To explore the changes of Bax/Bcl-2 in Cinobufotalin induced apoptosis in human breast cancer cell line MCF-7. Methods Human breast cancer cell line MCF-7 was exposed to Cinobufotal- in in different concentrations respectively. Each concentration group was exposed for 24 h , 48 h and 72 h. MTT assay was performed to detect the proliferation of the cells. The morphological changes of the cells were ob- served with the aid of inverted microscope. Apoptosis index was determined by Terminal deoxynucleotidyl trans ferase（TdT）-mediated dUTP nick end labeling （TUNEL） assay. Western blot analysis was used to test Bax/Bcl-2 expression. Bax distribution was assessed by immunofluorescence staining. Results Cinobufotalin inhibited the proliferation of MCF-7 cells. The percentage of apoptotic cells treated with Cinobufotalin increased as com pared to untreated cells. Western blotting revealed up-regulation of the Bax However, Cinobufotalin did not affect expression of the Bcl-2 protein. Bax was evenly distributed in cytoplasm in normal MCF-7 ceils, In constrast, Bax was translocated from cyto-plasm to mitochondria after Cinobufotalin treatment. Conclusions Cinobufotalin-induced apoptosis in MCF-7 ceils was accompanied with an up-expression of Bax protein and a mitochondrial translocation of the protein.protein.
出处
《中国中西医结合外科杂志》
CAS
2012年第6期580-583,共4页
Chinese Journal of Surgery of Integrated Traditional and Western Medicine
