【摘要】 目的探讨神经酰胺（cer）对人膀胱癌细胞的凋亡诱导作用及其机制。方法不同浓度C₂-神经酰胺（C₂-cer）作用于膀胱癌T24细胞,噻唑蓝（MTT）比色法检测细胞存活率,吖啶橙（AO）荧光染色、流式细胞术（FCM）检测细胞凋亡,并对半胱天冬酶（Caspase）-3活性、bcl-2蛋白含量和细胞色素C在细胞内的分布变化进行检测。结果T24细胞存活率与C₂-cer浓度呈负相关（r=-0．973,P＜0．01）。对照组和5、10、20、40μmol/L的C₂-cer处理组凋亡率分别为2．95％、9．18％、14．23％、29．12％、48．46％,C₂-cer处理组凋亡率显著高于对照组（P＜0．01）。Caspase-3活性随C₂-cer浓度增加而增加,与对照组比较差异有统计学意义（P＜0．05）。在C₂-cer作用下,T24细胞内bcl-2蛋白含量下降,细胞色素C向细胞质释放。结论降低bcl-2蛋白表达水平,从而促进线粒体细胞色素C释放和Caspase-3激活可能是C₂-cer诱导膀胱癌细胞凋亡的重要机制。更多还原

【Abstract】 Objective To study the apoptosis inducing effect and its mechanism of ceramide to human bladder cancer cells.Methods Different concentrations of C₂-ceramide（C₂-cer）were applied to human bladder cancer T24 cells,then MTT method was used to assay survival rates,fluorescent staining with acridine orange（AO）and flow cytometer（FCM）were used to detect cell apoptosis.Changes of Caspase-3 activity,bcl-2 protein and the intracellular distribution of cytochrome C were also detected.Re- sults The survival rate of T24 cells showed a negative correlation to the concentrations of C₂-cer（r= -0.973,P＜0.01）.The apoptotic rate in control group and the groups of 5,10,20,40μmol/L of C₂- cer was 2.95%,9.18%,14.23%,29.12% and 48.46%,respectively.The apoptotic rate in different concentrations of C₂-cer was higher than that of the control group（P＜0.01）.The activities of Caspase- 3 were increased with the concentrations of C₂-cer（P＜0.05 or P＜0.01）.During the apoptosis of T24 cells induced by C₂-cer,the amount of bcl-2 protein was decreased and cytochrome C was released from mitochondria.Conclusion C₂-cer may induce apoptosis of human bladder cancer T24 cells by decreasing the amount of bcl-2 protein,promoting the release of cytochrome C and the activation of Caspase-3.更多还原