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目的探讨平阳霉素抑制人脐静脉内皮细胞系ECV304的量效和时效关系及其相关机制。方法用不同浓度的平阳霉素作用于ECV304细胞24、48、72 h,MTT法观察细胞增殖抑制情况,电镜及DNA电泳实验分析细胞凋亡情况,流式细胞术分析细胞凋亡率、细胞周期及Fas、Bcl-2蛋白的表达。结果MTT法显示不同浓度平阳霉素组间细胞生长抑制率有显著差异,浓度越高、作用时间越长,抗增殖效应越明显;流式细胞术显示细胞凋亡率随药物浓度和作用时间增加而升高,平阳霉素各浓度作用24 h后,G2-M期百分率增高,S期百分率降低,G0-G1期百分率无明显变化;Fas表达随药物浓度增加而递增,Bcl-2表达随药物浓度增加而递减。结论平阳霉素能明显抑制ECV304细胞生长,并具有浓度和时间依赖性;平阳霉素可能是通过上调Fas蛋白并下调Bcl-2蛋白的表达,从而诱导细胞在G2-M期发生凋亡。
Objective To investigate the dose-response and time-effect relationship of pingyangmycin on human umbilical vein endothelial cell line ECV304 and its related mechanism. Methods ECV304 cells were treated with different concentrations of pingyangmycin for 24,48,72 h. MTT assay was used to observe the inhibition of cell proliferation. Cell apoptosis was analyzed by electron microscopy and DNA electrophoresis. Flow cytometry was used to analyze the apoptosis rate, cell cycle And Fas, Bcl-2 protein expression. Results MTT assay showed that there was a significant difference in the rate of cell growth inhibition between different concentrations of pingyangmycin. The higher the concentration and the longer the duration of action, the more obvious the anti-proliferative effect was. Flow cytometry showed that the apoptosis rate increased with the concentration and duration of action The percentages of G2-M phase increased, the percentage of S phase decreased, while the percentage of G0-G1 phase did not change significantly. The expression of Fas increased with the increase of drug concentration, and the expression of Bcl-2 increased with the increase of drug concentration Concentration increases and decreases. Conclusion Pingyangmycin can significantly inhibit the growth of ECV304 cells in a concentration-and time-dependent manner. Pingyangmycin may induce apoptosis in G2-M phase by up-regulating Fas protein and down-regulating Bcl-2 protein expression.