Silencing integrin-linked kinase by small interfering RNA inhibits metastasis and invasion of human EJ bladder cancer cells

朱军; 高娟; 李红彦; 潘湘阳; 陈俊霞

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Silencing integrin-linked kinase by small interfering RNA inhibits metastasis and invasion of human EJ bladder cancer cells

更新时间：2026-01-09

- Silencing integrin-linked kinase by small interfering RNA inhibits metastasis and invasion of human EJ bladder cancer cells
- Journal of Army Medical University Vol. 33, Issue 4, Pages: 360-364(2011)
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  CLC： R394-33;R73-362;R737.14
- Published：2011
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朱军,高娟,李红彦,潘湘阳,陈俊霞.特异性ILK-siRNA对膀胱癌EJ细胞侵袭和迁移的影响[J].陆军军医大学学报(原第三军医大学学报),2011,33(04):360-364.

　Zhu Jun,Gao Juan,Li Hongyan,Pan Xiangyang,Chen Junxia.Silencing integrin-linked kinase by small interfering RNA inhibits metastasis and invasion of human EJ bladder cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(04):360-364.
朱军,高娟,李红彦,潘湘阳,陈俊霞.特异性ILK-siRNA对膀胱癌EJ细胞侵袭和迁移的影响[J].陆军军医大学学报(原第三军医大学学报),2011,33(04):360-364. DOI：

　Zhu Jun,Gao Juan,Li Hongyan,Pan Xiangyang,Chen Junxia.Silencing integrin-linked kinase by small interfering RNA inhibits metastasis and invasion of human EJ bladder cancer cells[J].J Amry Med Univ (J Third Mil Med Univ),2011,33(04):360-364. DOI：

摘要

目的设计合成有效的ILK-siRNA

并检测其对EJ细胞侵袭和迁移的影响。方法构建针对ILK基因mRNA 的2个特异性siRNA表达载体pGenSil-1 siRNA1、pGensil-1 siRNA2和1个阴性对照载体pGenSil-1 siRNA3。稳定转染膀胱癌EJ细胞后，通过RT-PCR检测EJ细胞中ILK mRNA水平的表达，并通过Western blot和细胞免疫荧光检测ILK蛋白水平的表达；观察EJ细胞形态变化；检测EJ细胞迁移、黏附和侵袭能力及基质金属蛋白酶的变化。结果重组质粒构建成功。干扰组细胞ILK mRNA及蛋白的表达分别较3个对照组（未转染组、转空质粒组和转阴性对照质粒组）比较，呈显著性下降（P0.05）；HE染色结果显示，细胞铺展良好，核质比减小；转染干扰质粒的2个实验组细胞的黏附能力相对于对照组分别增加87%和76%，黏附能力显著升高，迁移及侵袭能力明显降低（P0.05）；与3个对照组比较

转染pGenSil-1 siRNA1与pGensil-1 siRNA2质粒到EJ细胞后

MMP-2及MMP-9的表达水平明显降低（P0.05）。结论成功构建的干扰质粒能抑制ILK基因及蛋白水平的表达，显著抑制细胞的生长、侵袭及迁移能力。

Abstract

Objective To investigate the effect of the specific siRNA of integrin-linked kinase (ILK) on the migration and invasion of human EJ bladder cancer cells. Methods Two specific siRNA expression vectors and one none homology control vector targeting the objective gene was constructed. Then the vectors were transfected into EJ cells respectively. RNAi efficiency on the expression of ILK was evaluated by RT-PCR

Western blotting and immunofluorescence staining. The morphological change

cell adhesion

migration and invasion ability

and the expressions of MMP2 and MMP9 of the transfected cells were detected. Results Enzyme digestion and DNA sequencing verified the constructed vectors were correct. In the 2 specific siRNA treatment groups

the expression of ILK was obviously inhibited

the cells spread well

the nuclear mass ratio was lower than control

the cell adhesion ability was increased

the cell migration and metastasis ability were decreased

and the expression of MMP2 and MMP9 obviously decreased. Conclusion The specific siRNA vectors of ILK are successfully constructed. The vectors inhibit ILK expression

and decrease cell migration and metastasis ability significantly in human EJ bladder cancer cells.

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