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

doi:10.13481/j.1671-587x.20150510

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (05): 937-940.doi: 10.13481/j.1671-587x.20150510

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

尹健¹, 张斯棋², 张钰³, 李然伟⁴

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

收稿日期:2015-03-23 出版日期:2015-09-28 发布日期:2015-09-29
通讯作者: 李然伟,教授,硕士研究生导师(Tel:0431-88796612,E-mail:ranwei1968@yahoo.com) E-mail:ranwei1968@yahoo.com
作者简介:尹健(1980-),男,吉林省长春市人,主治医师,医学硕士,主要从事血管生长因子与肿瘤关系方面的研究。
基金资助:
吉林省科技厅自然科学基金资助课题(200905139)

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

YIN Jian¹, ZHANG Siqi², ZHANG Yu³, LI Ranwei⁴

1. Department of Vascular Surgery, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
2. Department of Nephrology, China-Japan Union Hospital, Jilin University, Changchun 130033, China;
3. Department of Pathophysiology, School of Basic Medical Sciences, Jilin University, Changchun 130021, China;
4. Department of Urinary Surgery, Second Hospital, Jilin University, Changchun 130041, China

Received:2015-03-23 Online:2015-09-28 Published:2015-09-29

摘要/Abstract

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pcDNA-VEGF165b;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48 h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48 h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cleaved-caspase3/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。

关键词: 膀胱肿瘤, 血管内皮生长因子, 顺铂

Abstract:

Objective To observe the effects of exogenous expression of mutamer of vascular endothelial growth factor VEGF165b on the damage of human bladder cancer T24 cells induced by cisplatin(DDP),and to discuss its mechanism. Methods The VEGF165b expression vector pcDNA-VEGF165b was constructed.According to the processing method,T24 cells were divided into control group,VEGF165b group (transfected with VEGF165b expression vector),DDP group (treated with DDP),DDP+VEGF165b group (DDP plus transfected with VEGF165b expression 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 VEGF165b group at 24 and 48 h had no significant changes;compared with DDP group, the cell viability at 24 and 48 h in 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 T24 cells 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 VEGF165b+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 VEGF165b group (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 VEGF165b+DDP group (41.3+6.9) was increased (P<0.05). Conclusion The expression of exogenous VEGF165b can 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.

Key words: bladder neoplasms, vascular endothelial growth factor, cisplatin

中图分类号:

R737.14

尹健, 张斯棋, 张钰, 李然伟. 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[J]. 吉林大学学报(医学版), 2015, 41(05): 937-940.

YIN Jian, ZHANG Siqi, ZHANG Yu, LI Ranwei. Effect of exogenous expression of VEGF165b on injury of human bladder cancer T24 cells induced by cisplatin[J]. Journal of Jilin University Medicine Edition, 2015, 41(05): 937-940.

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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

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