【摘要】 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。更多还原

【Abstract】 Objective To investigate the concentration-effect and time-response effect relationships of Pingyangmycin(PYM) against human umbilical vein endothelial cell line ECV304 and the possible mechanism.Methods PYM was used in medium to culture ECV304 cells,and the concentration-effect and time-response effect relationships were evaluated by MTT assay.Apoptosis was observed by using transmission electron microscopy(TEM) and analyzed by electrophoresis of DNA.The apoptotic rate,cell cycle and the expression of Fas and Bcl-2 were analyzed by flow cytometry.Results The growth of ECV304 cells were obviously inhibited,and its inhibit rate increased with increase of PYM concentration and prolongation of culture time.Apoptotic rate increased with increase of PYM concentration and prolongation of culture time.The percentage of G2-M phase cells increased while the S phase decreased,but the percentage of G0-G1 phase cells remained unchanged after treated with Pingyangcin for 24h.The expression of Fas increased with increase of PYM concentration while the expression of Bcl-2 decreased.Conclusions PYM can effectively inhibit ECV304 cells in vitro with concentration-effect and time-response effect relationships.The mechanism probably is that PYM induce ECV304 cell apoptosis in the cell cycle of G2-M phase through the up-regulation of Fas and the down-regulation of Bcl-2.更多还原