【摘要】 目的观察丝裂霉素C（MMC）与C2-神经酰胺（C₂-cer）联合应用对人膀胱癌细胞的作用效果,并探讨其机制。方法不同浓度MMC与C₂-cer单独及联合作用于人膀胱癌BIU-87细胞后,应用噻唑蓝（MTT）比色法检测细胞生长抑制率,计算合用指数（CI）,流式细胞仪（FCM）检测BIU-87细胞凋亡率,吖啶橙（AO）荧光染色观察凋亡形态学变化,Western blot检测细胞色素C在细胞内分布变化,并检测Caspase-3活性改变。结果单独应用时MMC与C₂-cer的中效浓度分别是159和28μmol/L,联合用药时下降为55和11μmol/L,CI=0．74。MMC与G₂-cer单独及联合应用均可导致BIU-87细胞出现凋亡的形态学变化。两种药物联合应用时的凋亡率高于各自单用（P＜0．05）。线粒体细胞色素C含量在MMC与C₂-cer单独及联合应用时均较对照组减少,联合用药时减少最为明显,细胞质内细胞色素C含量在联合用药时增加也最为明显（P＜0．05）。Caspase-3活性在MMC与C₂-cer单独及联合应用时均较对照组升高,联合用药时升高最为明显（P＜0．05）。结论MMC与C₂-cer联合应用可以通过共同诱导细胞凋亡,协同抑制膀胱癌细胞生长。线粒体细胞色素C释放和Caspase-3活性变化可能发挥重要作用。更多还原

【Abstract】 Objective To evaluate the therapeutic effectiveness of combined use of mitomycin C （MMC）with C₂-ceramide（C₂-cer）on human bladder cancer cells and the mechanism.Methods Different concentratious of MMC and C₂-cer were applied,alone or together,to human bladder cancer BIU-87 cells.MTT method was used to assay the effect of synergetic inhibition to cell growth by calculating concentration index （CI）.Flow eytometer（FCM）and fluorescent staining with acridine orange（AO）were used to detect cell apop- tosis.Western blot was used to detect the distribution changes of intracellular cytochrome C.At the same time, the changes of Caspase-3 activity were detected.Results The IC50 of MMC and C₂-cer was 159 and 28μmol/ L respectively when they were used alone,while that was 55 and 11μmol/L when they were combined.Combi- nation index（CI）was 0.74.Call apoptosis could be induced no matter MMC and C₂-cer were used alone or to- gether.The apoptosis rate was higher the group of MMC+C₂-cer than that in the group of MMC or C₂-cer alone.（P＜0.05）.The amounts of cytochrome C in mitochondria were decreased in the group of MMC+C₂- cer and group of MMC or C₂-cer used alone than in the control group,most obviously in the group of combined use.The amounts of cytochrome C in cytosol were also increased most obviously when two drugs were combined （P＜0.05）.The Caspase-3 activity of BIU-87 cells was increased in the group of MMC+C₂-cer and group of MMC or C₂-cer alone than in the control group,most obviously in the group of MMC+C₂-cer（P＜0.05）. Conclusion The combined use of MMC and C₂-cer can promote cell apoptosis and inhibit the growth of bladder cancer cells synergistically.The release of cytochrome C from mitochondria to cytosol and the changes of Cas- pase-3 activity may play a critical role during this process.更多还原