丝裂霉素C与C_2-神经酰胺联合应用治疗膀胱癌  

Experimental study on the therapeutic effectiveness of combined use of mitomycin C with C_2-ceramide on human bladder cancer cells

作　　者：时京[1] 孔垂泽[1] 孙志熙[1] 王侠[1] 罗阳[2] 刘奔[1] 张莹[3] 刘博[1] 

机构地区：[1]中国医科大学附属第一医院泌尿外科,沈阳110001 [2]中国医科大学细胞生物学卫生部重点实验室医学基因组学教研室 [3]中国医科大学实验技术中心

出　　处：《中华实验外科杂志》2006年第10期1162-1164,i0001,共4页Chinese Journal of Experimental Surgery

基　　金：辽宁省自然科学基金(20042082)

摘　　要：目的观察丝裂霉素C(MMC)与C2-神经酰胺(C_2-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C_2-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C_2-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G_2-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C_2-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C_2-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C_2-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。Objective To evaluate the therapeutic effectiveness of combined use of mitomycin C （MMC） with C2-ceramide （C2-cer） on human bladder cancer ceils and the mechanism. Methods Different concentrations of MMC and C2-cer were applied,alone or together, to human bladder cancer BIU-87 ceils. MTT method was used to assay the effect of synergetic inhibition to cell growth by calculating concentration index （CI）. Flow cytometer （FCM） and fluorescent staining with acridine orange （AO） were used to detect cell apoptosis. 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 28 μmol/ L respectivdy when they were used alone,while that was 55 and 11 μmol/L when they were combined. Combination index （CI） was O. 74. Cell apoptosis could be induced no matter MMC and C2-cer were used alone or together. 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 MMCor 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 Caspase-3 activity may play a critical role during this process.

关 键 词：膀胱肿瘤 丝裂霉素C 神经酰胺 脱噬作用 

分 类 号：R737.14[医药卫生—肿瘤]