外源性表达VEGF165b对人膀胱癌T24细胞侵袭力的影响

doi:10.13481/j.1671-587x.20150113

吉林大学学报(医学版) ›› 2015, Vol. 41 ›› Issue (01): 64-68.doi: 10.13481/j.1671-587x.20150113

外源性表达VEGF165b对人膀胱癌T24细胞侵袭力的影响

尹健¹, 张斯棋², 冯树强³, 张金³, 李然伟³

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. 吉林大学第二医院泌尿外科, 吉林长春 130041

收稿日期:2014-08-14 发布日期:2015-01-30
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 invasion ability of human bladder cancer T24 Cells

YIN Jian¹, ZHANG Siqi², FENG Shuqiang³, ZHANG Jin³, LI Ranwei³

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 Urinary Surgery, Second Hospital, Jilin University, Changchun 130041, China

Received:2014-08-14 Published:2015-01-30

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T24细胞生存率、迁移率和侵袭力的作用,探讨VEGF165b对人膀胱癌细胞生物学特性的影响。方法: 构建VEGF165b表达载体pcDNA-VEGF165b;T24细胞根据处理因素不同分为T24细胞组(正常对照)、pcDNA3.0组(转染pcDNA3.0表达载体)和pcDNA-VEGF165b组(转染pcDNA-VEGF165b表达载体)。MTT法检测T24细胞生存率, Western blotting法检测T24细胞中VEGF165b、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)蛋白表达水平,Transwell小室法检测T24细胞迁移率和穿膜细胞数。结果: 与T24细胞组比较,pcDNA3.0组T24细胞中VEGF165b、MMP-2和MMP-9蛋白表达水平无明显变化(P>0.05)。与T24细胞组和pcDNA3.0组比较,pcDNA-VEGF165b组细胞中VEGF165b蛋白表过水平明显升高(P<0.05),MMP-2和MMP-9蛋白表达水平明显降低(P<0.05)。T24细胞组、pcDNA3.0组和pcDNA-VEGF165b组细胞存活率比较差异无统计学意义(P>0.05)。与T24细胞组细胞迁移率(100%)比较,pcDNA3.0组细胞迁移率(97.2%±7.8%)无明显变化(P>0.05),pcDNA-VEGF165b组细胞迁移率(63.5%±10.4%)明显降低(P<0.05)。细胞侵袭力实验,与T24细胞组穿膜细胞数(70.8±8.5)比较,pcDNA3.0组穿膜细胞数(66.3±11.2)无明显变化(P>0.05),pcDNA-VEGF165b组穿膜细胞数(23.5±7.3)明显降低(P<0.05)。结论: 外源性表达VEGF165b对T24细胞增殖无明显影响,但可明显降低其迁移力和侵袭力。

关键词: 膀胱肿瘤, 血管内皮生长因子165b, 肿瘤浸润

Abstract:

Objective To observe the effects of exogenous expression of VEGF165b on the viability, migrationrate and invasion ability of human bladder cancer T24 cells, and to explore the infulence of VEGF165b in the bilogical features of human bladder cancer cells. Methods The VEGF165b expression vector pcNDA-VEGF165b was constructed.The T24 cells were divided into T24 cells group (control), pcDNA3.0 group (transfected by pcDNA3.0 plasmid) and pcDNA-VEGF165b group (transfected by pcDNA-VEGF165b plasmid).The viability of T24 cells was detected by MTT assay;Western blotting method was used to determine the expressions of VEGF165b, matrix metalloproteinase-2(MMP-2) and matrix metalloproteinase-9(MMP-9) proteins.Transwell assay was used to detect the rate of cell migration and invasive ability. Results Compared with T24 cells group, the expression levels of VEGF165b, MMP-2 and MMP-9 proteins in the T24 cells in pcDNA3.0 grouphad no significant differences (P>0.05). Compared with T24 cells group or pcDNA3.0 group, the expression level of VEGF165b protein in the T24 cells in pcDNA-VEGF165b group was significantly increased(P<0.05), and the expression levels of MMP-2 and MMP-9 proteins were significantly decreased(P<0.05).The cell viabilities of T24 cells had no significant difference between three groups (P>0.05).Compared with T24 cells group (migration rate 100%), the migration rate of the T24 cells in pcDNA3.0 group (97.2%±7.8%) had no significant difference(P>0.05), but the migration rate in pcDNA-VEGF165b group was decreased significantly (63.5%±10.4%)(P<0.05). Compared with T24 cells group (70.8±8.5), the number of transferred T24 cells in pcDNA3.0 group (66.3±11.2) had no significant difference(P>0.05), but in pcDNA-VEGF165b group (23.5±7.3) it was decreased significantly (P<0.05). Conclusion Exogenous expression of VEGF165b in the T24 cells has no significant effect on the proliferation of T24 cells, but it can reduce the migration and invasion abilities significantly.

Key words: bladder neoplsms, vascular endothelial growth factor 165b, tumor invasion

中图分类号:

R737.14

尹健, 张斯棋, 冯树强, 张金, 李然伟. 外源性表达VEGF165b对人膀胱癌T24细胞侵袭力的影响[J]. 吉林大学学报(医学版), 2015, 41(01): 64-68.

YIN Jian, ZHANG Siqi, FENG Shuqiang, ZHANG Jin, LI Ranwei. Effect of exogenous expression of VEGF165b on invasion ability of human bladder cancer T24 Cells[J]. Journal of Jilin University Medicine Edition, 2015, 41(01): 64-68.

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外源性表达VEGF165b对人膀胱癌T24细胞侵袭力的影响

Effect of exogenous expression of VEGF165b on invasion ability of human bladder cancer T24 Cells

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