重组人程序化细胞死亡5蛋白对顺铂诱导膀胱癌细胞凋亡的影响  

Effect of recombinant human programmed cell death 5 protein on cell apoptosis of human bladder cancer cell line BIU87 induced by cisplatin and its mechanism

作　　者：严泽军[1] 夏术阶[2] 蒋军辉[1] 谢国海[1] 钱君海[1] 蒋照辉[1] 周成[1] 程跃[1] 

机构地区：[1]宁波大学医学院附属宁波市第一医院泌尿外科,浙江315000 [2]上海交通大学附属第一人民医院泌尿外科

出　　处：《中华实验外科杂志》2015年第6期1220-1223,共4页Chinese Journal of Experimental Surgery

基　　金：中国博士后科学基金资助项目（2013M541524）;浙江省自然科学基金资助项目（LY12H05002）;浙江省医药卫生科技计划资助项目（2013KYA180）;宁波市自然科学基金资助项目（2013A610208）

摘　　要：目的 观察重组人程序化细胞死亡5（rhPDCD5）蛋白对顺铂（DDP）诱导人膀胱癌BIU87细胞凋亡的影响,并探讨其作用机制.方法 采用细胞计数试剂盒（CCK-8）法检测不同浓度的rhPDCD5蛋白和/或DDP对膀胱癌BIU87细胞抑制率的影响;Hoechst 33342染色观察细胞的形态变化;流式细胞术（FCW）检测细胞凋亡率;采用Western blot法检测细胞中含半胱氨酸的天冬氨酸蛋白水解酶-3（Caspase-3）和B细胞淋巴瘤/白血病-2（bcl-2）蛋白的表达.结果 CCK-8检测结果显示,rhPDCD5蛋白与DDP能有效抑制BIU87细胞的增殖.Hoechst 33342染色检测显示,rhPDCD5＋DDP联合用药组阳性细胞百分比为（31.62±2.33）％,显著高于单用rhPDCD5蛋白组的（9.61±0.42）％和单用DDP组的（14.12±0.24）％,差异有统计学意义（F=221.42,P＜0.01）.FCM检测结果显示,rhPDCD5＋ DDP联合用药组凋亡率为（58.74±3.48）％,与rhPDCD5组的（18.84±1.23）％和DDP组的（21.25±2.27）％比较,细胞凋亡率明显提高（F=228.93,P＜0.01）.Western blot结果显示,联合用药组较单用药组细胞凋亡相关蛋白Caspase-3表达明显增强（F=12.60,P＜0.01）,bcl-2表达显著减少（F=12.20,P＜0.01）.结论 rhPDCD5蛋白可直接作用于BIU87细胞使细胞凋亡,并对DDP诱导的BIU87细胞凋亡有明显促进作用.Objective To observe the effect of recombinant human programmed cell death 5 （rhPDCD5） protein on the apoptosis of human bladder cancer cell line BIU87 induced by cisplatin （DDP） and to investigate its mechanism.Methods Inhibition rates of bladder cancer cell line BIU87 intervened by rhPDCD5 protein and/or DDP with different concentrations were tested by cell counting kit-8 （CCK-8）.The morphological changes of cells were detected by Hoechst 33342 staining.The cells apoptosis rates were examined by flow cytometry.The changes of expression of the cysteinyl aspartate specific proteinase-3 （Caspase-3） and B-cell lymphoma/Leukemia-2 （bcl-2） were measured by Western blotting analysis.Results CCK-8 showed that rhPDCD5 protein or/and DDP could inhibit the proliferation of BIU87 cells effectively.Hoechst 33342 staining showed that the positive cells rate of cells treated with rhPDCD5 protein and DDP （31.62 ± 2.33）％ was obviously higher than that only treated with rhPDCD5 protein （9.61 ± 0.42） ％ or DDP （14.12 ± 0.24） ％,the difference was significant （F =221.42,P ＜ 0.01）.The flow cytometry （FCM） showed that the apoptosis rate of cells treated with rhPDCD5 protein and DDP （58.74 ± 3.48）％ was obviously increased than that only treated with rhPDCD5 protein （18.84 ± 1.23） ％ or DDP （21.25 ± 2.27） ％,the difference was significant （F =228.93,P ＜ 0.01）.Compared with the group only treated with rhPDCD5 protein or DDP,the expression of Caspase-3 of the group treated with rhPDCD5 protein and DDP was significantly up-regulated （F =12.60,P ＜ 0.01） and the expression of bcl-2 was significantly down-regulated （F =12.20,P ＜ 0.01）.Conclusion rhPDCD5 protein can induce the apoptosis of BIU87 cells,and accelerate the apoptosis of BIU87 cells induced by DDP.rhPDCD5 protein may reduce the expression of bcl-2 protein and increase activation of Caspase-3 to play its role in promoting apoptosis.

关 键 词：程序化细胞死亡分子5 顺铂 膀胱肿瘤 细胞凋亡 

分 类 号：R737.14[医药卫生—肿瘤]