外源性表达VEGF165b对顺铂诱导人膀胱癌T24细胞损伤的影响及其机制  

Effect of exogenous expression of VEGF165b on injury of human bladder cancer T24 cells induced by cisplatin

作　　者：尹健[1] 张斯棋[2] 张钰[3] 李然伟[4] 

机构地区：[1]吉林大学中日联谊医院血管外科,吉林长春130033 [2]吉林大学中日联谊医院肾病内科,吉林长春130033 [3]吉林大学基础医学院病理生理学系,吉林长春130021 [4]吉林大学第二医院泌尿外科,吉林长春130041

出　　处：《吉林大学学报（医学版）》2015年第5期937-940,I0003,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅自然科学基金资助课题(200905139)

摘　　要：目的:观察人膀胱癌T24细胞中外源性表达血管内皮生长因子(VEGF)变构体VEGF165b对顺铂(DDP)诱导人膀胱癌T24细胞损伤的作用,并探讨相关机制。方法:构建VEGF165b表达载体pcDNAVEGF165b;T24细胞分为对照组、VEGF165b组(转染VEGF165b表达载体)、DDP组和VEGF165b+DDP组(DDP联合转染VEGF165b表达载体)。MTT法检测各组细胞存活率;Western blotting法检测各组T24细胞中凋亡相关蛋白Bcl-2、Bax和caspase3表达;TUNEL法检测细胞凋亡数。结果:MTT法检测,与对照组比较,VEGF165b组24和48h时T24细胞存活率无明显变化(P>0.05);与DDP组比较,VEGF165b+DDP组24和48h时细胞存活率均明显降低(P<0.05)。Western blotting法检测,与对照组比较,DDP组细胞中cleavedcaspase3/pro-caspase3比值升高(P<0.05),Bcl-2/Bax比值降低(P<0.05);与DDP组比较,VEGF165b+DDP组细胞中cleaved-caspase3/pro-caspase3比值升高(P<0.05),Bcl-2/Bax比值降低(P<0.05)。TUNEL法检测,与对照组(5.1±2.7)和VEGF165b组(7.4±3.2)比较,DDP组的凋亡细胞数(26.3±8.2)的凋亡细胞数增加(P<0.05)。与DDP组比较,VEGF165b+DDP组的凋亡细胞数(41.3±6.9)增加(P<0.05)。结论:外源性表达VEGF165b能通过增加细胞凋亡促进DDP对T24细胞的损伤,其机制与VEGF信号通路受抑制有关联。Objective To observe the effects of exogenous expression of mutamer of vascular endothelial growth factor VEGF165 bon the damage of human bladder cancer T24 cells induced by cisplatin（DDP）,and to discuss its mechanism.Methods The VEGF165 bexpression vector pcDNA-VEGF165 bwas constructed.According to the processing method,T24 cells were divided into control group,VEGF165bgroup（transfected with VEGF165 bexpression vector）,DDP group（treated with DDP）,DDP＋VEGF165bgroup（DDP plus transfected with VEGF165 bexpression vector）.MTT method was used to detected the cell viability;Western blotting method was used to detect the expression of apoptotic proteins Bcl-2,Bax,and caspase3;TUNEL method was used to detect the number of apoptotic cells.Results The MTT results showed that compared with control group,the cell viability in VEGF165 bgroup at 24 and 48hhad no significant changes;compared with DDP group,the cell viability at 24 and 48hin VEGF165b＋ DDP group were significantly decreased（P〈0.05）.The Western blotting results showed that compared with control group,the value of cleaved-caspase3/pro-caspase3 in T24cells in DDP group was increased（P〈0.05）,and the value of Bcl-2/Bax was decreased（P〈0.05）;compared with DDP group,the value of cleaved-caspase3/pro-caspase3 in cells in VEGF165 b ＋ DDP group was increased（P〈0.05）,and the value of Bcl-2/Bax was decreased（P〈0.05）.The TUNEL results showed that compared with control group（5.1±2.7）and VEGF165bgroup（7.4±3.2）,the number of apoptotic cells in DDP group（26.3±8.2）was increased（P〈0.05）.Compared with DDP group,the number of apoptotic cells in VEGF165 b ＋ DDP group（41.3 ＋6.9）was increased（P〈0.05）.Conclusion The expression of exogenous VEGF165 bcan promote the damage of DDP on T24 cells by increasing the apoptosis of T24 cells,and its mechanism is related to the inhibition of VEGF siginal passway.

关 键 词：膀胱肿瘤 血管内皮生长因子 顺铂 

分 类 号：R737.14[医药卫生—肿瘤]