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目的观察硫利达嗪对人乳腺癌细胞MDA-MB-231和MCF-7的凋亡作用,并探讨其机制。方法采用MTT法测定硫利达嗪对细胞的抑制作用,计算半数抑制浓度(IC50);流式细胞术检测硫利达嗪对细胞周期分布和凋亡的影响;比色法测定药物对细胞Caspase-3活性的影响;Western blot法检测凋亡调节蛋白Bcl-2、Bax表达的变化。结果硫利达嗪作用24 h后,MDA-MB-231、MCF-7细胞增殖明显呈剂量依赖性抑制,其IC50为18、22μmol/L。流式细胞术结果显示,随着加入硫利达嗪浓度的提高,MDA-MB-231、MCF-7细胞均发生不同程度的G0/G1期阻滞、细胞凋亡程度增加及伴随胞内Caspase-3活性增加。各实验组与对照组比较,差异均有统计学意义(P<0.01)。Western blot法结果显示随着药物浓度的增加,抗凋亡蛋白Bcl-2表达下调、促凋亡蛋白Bax表达明显上调,各实验组与对照组相比,差异有统计学意义(P<0.01)。结论硫利达嗪对乳腺癌细胞MDA-MB-231、MCF-7有显著的增殖抑制作用且可显著诱导肿瘤细胞发生G0/G1期阻滞及凋亡,伴随Caspase-3活性的上调,其毒性机制可能与肿瘤细胞内抗凋亡蛋白Bcl-2下调、Bax上调有关。
Objective To observe the apoptosis of human breast cancer cells MDA-MB-231 and MCF-7 induced by thioridazine and explore its mechanism. Methods MTT assay was used to determine the inhibitory effect of thioridazine on IC50. The IC50 was calculated. The effect of thioridazine on cell cycle distribution and apoptosis was analyzed by flow cytometry. -3 activity. Western blot was used to detect the expression of Bcl-2 and Bax. Results After treated with thioridazine for 24 h, the proliferation of MDA-MB-231 and MCF-7 cells significantly inhibited in a dose-dependent manner with IC50 of 18 and 22 μmol / L. Flow cytometry showed that with the increase of the concentration of thioridazine, the G0 / G1 phase arrest and the degree of apoptosis were increased in MDA-MB-231 and MCF-7 cells with the increase of intracellular Caspase -3 activity increased. The experimental group and control group, the difference was statistically significant (P <0.01). The results of Western blot showed that with the increase of drug concentration, the expression of anti-apoptotic protein Bcl-2 was down-regulated and the expression of pro-apoptotic protein Bax was up-regulated. There was significant difference between the experimental group and the control group (P <0.01) . Conclusions Thioridazine can significantly inhibit the proliferation of breast cancer cells MDA-MB-231 and MCF-7 and induce G0 / G1 phase arrest and apoptosis in tumor cells with an upregulation of Caspase-3 activity Toxicity may be related to down-regulation of Bcl-2 and up-regulation of Bax in tumor cells.