MMC诱导EJ细胞凋亡和Fas/FasL、Caspase-3基因表达

MMC-Induced Apoptosis and Expression of Fas /FasL and Caspase-3 genes in EJ Cells

目的:研究丝裂霉素(MMC)在体外诱导膀胱癌细胞EJ凋亡和凋亡相关基因Fas/FasL、Caspase-3表达的关系。方法:以低剂量MMC(0.100mg/ml)诱导EJ细胞凋亡;以TUNEL法和FCM研究EJ细胞凋亡、细胞周期分布及Caspase-3抗体对抗MMC诱导凋亡;以免疫组化检测Fas/FasL、Caspase-3蛋白表达。结果:低剂量MMC处理的EJ细胞出现明显的凋亡形态特征,凋亡峰和细胞生长阻滞均发生于G0/G1期,Fas/FasL、Caspase-3蛋白呈上调表达,Caspase-3抗体能特异性阻断MMC诱导EJ细胞凋亡。结论:低剂量MMC能够诱导EJ细胞凋亡和增加Fas/FasL、Caspase-3基因表达,且与启动Caspase凋亡信号传导有关。

Objective:To study the relationship between apoptosis of EJ cells induced by mitomycin C(MMC)and the expression of Fas /FasL,caspase-3genes.Methods:Low dose MMC was used to induce the apoptosis of EJ cells.TUNEL methiod and FCM method were used for detecting apoptosis of EJ cells,analysis of cell cycle,the influence of Caspase-3anti-body against the MMC inducing apoptosis and the expression of Fas /FasL,caspase-3protein de-tected by immunohistochemical method.Results:The EJ cells treated by low dose MMC demonstrated prominent morphological features and the apoptic peak and retardation of cellular growth all occurred in G 0 /G 1 phase,and the expression of Fas /FasL and caspase-3protein was up-regulated.The antibody The caspase-3antibody could resist the EJ cell apoptosis induced by MMC specifically.Conclusion:Low dose MMC not only can induce the apoptosis of EJ cells and increase the expression of Fas /FasL and caspase-3genes,but also is related to the start of conducting caspase apoptosis message.