过表达人RI抑制人膀胱癌T24细胞的侵袭、转移及EMT发生

姚雪; 熊冬梅; 舒静; 李林; 陈俊霞

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过表达人RI抑制人膀胱癌T24细胞的侵袭、转移及EMT发生

论著 | 更新时间：2026-01-09

- 过表达人RI抑制人膀胱癌T24细胞的侵袭、转移及EMT发生
- Over-expression of human ribonuclease inhibitor suppresses invasion, migration and EMT in human bladder cancer T24 cells
- 陆军军医大学学报 2012年34卷第23期页码：2378-2383
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  中图分类号： R394.2;R73-37;R737.14
- 纸质出版：2012
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姚雪,熊冬梅,舒静,李林,陈俊霞.过表达人RI抑制人膀胱癌T24细胞的侵袭、转移及EMT发生[J].陆军军医大学学报(原第三军医大学学报),2012,34(23):2378-2383.

　Yao Xue,Xiong Dongmei,Shu Jing,Li Lin,Chen Junxia.Over-expression of human ribonuclease inhibitor suppresses invasion, migration and EMT in human bladder cancer T24 cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(23):2378-2383.
姚雪,熊冬梅,舒静,李林,陈俊霞.过表达人RI抑制人膀胱癌T24细胞的侵袭、转移及EMT发生[J].陆军军医大学学报(原第三军医大学学报),2012,34(23):2378-2383. DOI：

　Yao Xue,Xiong Dongmei,Shu Jing,Li Lin,Chen Junxia.Over-expression of human ribonuclease inhibitor suppresses invasion, migration and EMT in human bladder cancer T24 cells[J].J Amry Med Univ (J Third Mil Med Univ),2012,34(23):2378-2383. DOI：

摘要

目的检测过表达人核糖核酸酶抑制因子(ribonuclease inhibitor，RI) 基因的质粒pIRES2-EGFP-RI对人膀胱癌T24细胞侵袭转移及上皮细胞间质转化（epithelial mesenchymal transition，EMT）现象的影响。方法将连有RI的质粒pIRES2-EGFP-RI稳定转染T24细胞后利用G418筛选出阳性克隆。根据转染的质粒不同分为3组：T24组（未转染组）、T24-0（转染pIRES2-EGFP质粒）和T24-RI组（转染pIRES2-EGFP-RI质粒）。RT-PCR检测T24细胞中RI的mRNA水平变化，细胞免疫荧光及Western blot检测RI蛋白的表达，利用HE染色观察其细胞形态的变化，Transwell方法检测细胞侵袭及转移能力的变化，Western blot检测侵袭转移及EMT相关蛋白Twist、MMP-9 和Snail及E-cadherin的表达，构建移植瘤模型，利用HE染色观测肺组织切片癌细胞转移情况，组织免疫化学方法检测移植瘤中RI、Twist、Snail 及E-cadherin的表达。结果 T24-RI组人RI的mRNA水平及蛋白表达显著升高(P0.05)；HE染色结果表明细胞铺展良好，核质比明显减小，细胞由间质形态向上皮形态转变；Transwell方法显示T24-RI组细胞的侵袭及转移能力较T24组、T24-0组明显下降；Western blot结果表明T24-RI组Twist、MMP-9 和Snail较T24组、T24-0组显著降低(P0.01)

E-cadherin较T24组、T24-0组显著升高(P0.01)；HE染色肺组织切片显示T24-RI组较其他两组癌细胞转移降低，组织免疫化学结果表明T24-RI组RI、E-cadherin 蛋白明显升高，而Twist、Snail显著降低。结论过表达人RI载体提高了RI蛋白的表达水平，并抑制了人膀胱癌T24细胞EMT的发生，从而抑制了膀胱癌T24细胞的侵袭、转移。

Abstract

Objective To observe the effect of over-expression of human ribonuclease inhibitor (RI) on the invasion

migration and epithelial mesenchymal transition (EMT) in huma

n bladder cancer T24 cells. Methods The vectors of pIRES2-EGFP (T24-0 group) and pIRES2-EGFP-RI (T24-RI group) were used to transfect human bladder cancer T24 cells

respectively

and the positive clones were screened by G418. The T24 cells without transfection were served as control (T24 group). The expression of RI gene at mRNA and protein levels was evaluated by RT-PCR

immunofluorescence and Western blot analysis. HE staining was used for detect the changes of cell morphology. The protein levels of twist

matrix metalloproteinase 9 (MMP-9)

snail and E-cadherin were determined by Western blot analysis. Tumor cells at 2×10

of the three groups were respectively injected subcutaneously into the back of BALB/c nude mice to establish the xenograft models. HE staining was performed to observe the EMT and metastasis of the T24 cells in lung tissues

and immunohistochemical assay was applied to detect the expression of RI

E-cadherin

twist and snail in the tumors. Results Compared with the T24 group and T24-0 group

the expression of RI was significantly higher in the T24-RI group (P0.05). The cells spread well

the nucleo-cytoplasmic ratio decreased

and EMT was observed. The cell invasion and migration abilities significantly reduced in the T24-RI group compared with the T24 group and T24-0 group. The expression of twist

MMP-9 and snail significantly decreased

but the expression of E-cadherin significantly increased in the T24-RI group compared with the T24 group and T24-0 group (P0.01). Cell invasion and metastasis reduced in the lung tissues of BALB/c nude mice receiving an injection of pIRES2-EGFP-RI-transfected T24 cells. The expression of RI and E-cadherin significantly increased

but the expression of twist and snail significantly decreased in the T24-RI group compared with the T24 group and T24-0 group. Conclusion Over-expression of RI can suppress EMT in bladder cancer T24 cells

and subsequently inhibit cell invasion and metastasis.

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