三氧化二砷诱导乳腺癌细胞凋亡以及对Survivin、Caspase-3蛋白表达的影响

Arsenic Trioxide Induced Apoptosis in MDA-MB-231 Cells and Its Influence on Expression of Survivin and Caspase-3 Proteins

背景与目的:探讨不同浓度As2O3在诱导人乳腺癌细胞MDA_MB_231凋亡过程中的作用,并研究其对Survivin和Caspase_3蛋白表达的影响。材料与方法:将3种不同浓度的As2O3(10.0、20.0、40.0μmol/L)与MDA_MB_231细胞共培养不同时间后,采用流式细胞术检测MDA_MB_231细胞凋亡;用免疫细胞化学技术及Westernblot检测Survivin和Caspase_3蛋白的表达。各实验均设相应的对照组。结果:经10.0、20.0、40.0μmol/LAs2O3作用后,MDA_MB_231细胞凋亡率分别为(28.89±2.47)%、(46.73±3.82)%和(56.44±4.16)%,细胞凋亡率在各浓度组间的差异均具有统计学意义(P均<0.01);免疫细胞化学染色显示Survivin蛋白表达减少,而Caspase_3蛋白被激活;Westernblot条带显示Survivin蛋白表达量降低,而Caspase_3蛋白则出现被激活的小片段。As2O3上述的3种作用均存在剂量依赖关系。结论:As2O3可能通过抑制Survivin蛋白活性,激活Caspase_3蛋白的表达而诱导MDA_MB_231细胞凋亡。

BACKGROUND AND AIM： To investigate the apoptosis in MDA-MB-231 cells induced by different concentrations of As2O3 and to study its influence on the activities of Survivin and Caspase-3 proteins. MATERIALS AND METHODS： The rate of apoptosis in human breast carcinoma MDA-MB-231 cells was detected by flow cytometry; the activities of Survivin and Caspase-3 proteins were examined by immunocytochemistry and Westem blot in untreated and As ：O rtreated MDA-MB-231 cells. REULTS： The percentages of the apoptosis in MDA-MB-231 cells were （28.89 ± 2.47）%,（46.73±3.82）% and （56.44±4.16）%, in cells treated for 24 hours with 10.0,20.0,40.0 μmol/L As2O3 respectively. As2O3 inhibited the expression of Survivin protein and promoted the expression of Caspase-3 protein. There were obvious dose-effect correlations between As2O3 and its functions. CONCLUSION： The results suggested that As2O3 could induce apoptosis of MDA-MB-231 cells via Survivin protein inhibition and Caspase-3 protein promotion.