Rce1在膀胱癌的表达及其对T24细胞凋亡的影响

曾强锋; 黄亮亮; 王德林; 何江; 吴文强; 何云锋; 李英; 麦力; 李建君; 刘俊男

doi:10.13406/j.cnki.cyxb.001006

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Rce1在膀胱癌的表达及其对T24细胞凋亡的影响

心、肺、泌尿疾病 | 更新时间：2024-01-17

- Rce1在膀胱癌的表达及其对T24细胞凋亡的影响
- Expression of Rce1 in bladder cancer and its influence on the apoptosis of T24 cells
- 重庆医科大学学报 2016年41卷第9期页码：897-903
- 作者机构：
  
  1. 重庆医科大学附属第一医院泌尿外科
  2. 重庆医科大学附属大学城医院消化神经中心
  3. 重庆市綦江区人民医院泌尿外科
  4. 重庆医科大学生命科学院
  5. 重庆医科大学生物化学与分子生物学肿瘤研究中心
- 作者简介：
- 基金信息：
  
  重庆市基础科学与前沿技术研究专项资助项目（编号:cstc2015jcyj B0228）;重庆市渝中区科技计划资助项目（编号:20150111）;重庆市卫生和计划生育委员会基金资助项目（编号:20143061）
- DOI：10.13406/j.cnki.cyxb.001006
  中图分类号： R737.14
- 纸质出版日期：2016，
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[1]曾强锋,黄亮亮,王德林,何江,吴文强,何云锋,李英,麦力,李建君,刘俊男.Rce1在膀胱癌的表达及其对T24细胞凋亡的影响[J].重庆医科大学学报,2016,41(09):897-903.

Zeng Qiangfeng, Huang Liangliang, Wang Delin, et al. Expression of Rce1 in bladder cancer and its influence on the apoptosis of T24 cells[J]. Journal of Chongqing Medical University, 2016,41(9):897-903.
[1]曾强锋,黄亮亮,王德林,何江,吴文强,何云锋,李英,麦力,李建君,刘俊男.Rce1在膀胱癌的表达及其对T24细胞凋亡的影响[J].重庆医科大学学报,2016,41(09):897-903. DOI： 10.13406/j.cnki.cyxb.001006.

Zeng Qiangfeng, Huang Liangliang, Wang Delin, et al. Expression of Rce1 in bladder cancer and its influence on the apoptosis of T24 cells[J]. Journal of Chongqing Medical University, 2016,41(9):897-903. DOI： 10.13406/j.cnki.cyxb.001006.

摘要

目的:探讨Rce 1（Ras and a-factor converting enzyme 1）在膀胱癌组织及对应癌旁组织中的表达差异及其意义;探究慢病毒介导sh RNA沉默Rce1基因对人膀胱癌细胞株T24细胞凋亡的影响。方法:采用q RT-PCR和Western blot技术检测Rce1在12例膀胱癌组织及对应癌旁组织中m RNA和蛋白表达情况

采用免疫组化技术检测76例膀胱癌组织及对应癌旁组织中Rce1表达情况;结合临床病理资料

分析Rce1表达与临床病理特征的关系。设计针对Rce1基因的sh RNA慢病毒干扰载体感染T24细胞

经嘌呤霉素筛选后建立稳定感染细胞系

实时荧光定量PCR、Western blot分别检测感染后各组细胞Rce1 m RNA和蛋白表达水平;应用流式细胞仪检测T24细胞凋亡改变情况。结果:q RT-PCR和Western blot显示在12对癌及癌旁组织中Rce1差异性表达

差异有统计学意义（P=0.00）;78例癌组织中Rce1阳性表达率为70.5%（55/78）

癌旁组织中阳性率为19.0%（4/21）

组间差异有统计学意义（P=0.000）;统计学分析表明

Rce1表达量与肿瘤组织学分级、T分期和肿瘤是否远处转移存在相关关系（P<0.05）;Kaplan-Meier生存分析显示

Rce1阳性表达与病人术后生存时间呈负相关关系（P=0.016）。与空白对照组和阴性对照组相比

感染sh RNA慢病毒干扰载体细胞组Rce1 m RNA和蛋白表达水平明显下调（P=0.000）

细胞凋亡明显增加。结论:Rce1在膀胱癌组织中表达高于癌旁组织

Rce1在膀胱癌的发生发展中可能发挥重要作用;慢病毒介导sh RNA沉默Rce1基因能有效下调Rce1在T24细胞中表达

并诱导细胞凋亡;Rce1有望成为膀胱癌临床诊治和预后的一个新指标。

Abstract

Objective:To investigate the expression and clinical significance of Ras and a-factor converting enzyme 1(Rce1)in tissues of bladder urothelial carcinoma(UHC)and pericarcinoma(PC)and to explore the effect of lentivirus-mediated sh RNA silencing Rce1 gene on apoptosis and cell cycle of human bladder cancer T24 cells. Methods:Immunohistochemical staining for Rce1 expression was performed in bladder urothelial carcinoma(UCB) and adjacent pericarcinoma tissues;the methods of q RT-PCR and Western blot were utilized to investigate m RNA/protein expression of Rce1 in UCB. Rce1(+)and Rce1(-)patients were compared in terms of p TNM grade and differentiation grade. Lentiviral interference vectors of sh RNA-Rce1 were constructed successfully and were transfected into T24 cells

then the T24 cells stably knockdown Rce1 gene expression were screened by puromycin. Expression of Rce1 m RNA and protein in T24 cells was detected by q RT-PCR and Western blot

respectively. Apoptosis of T24 cells was examined by flow cytometry. Results:The expressions of Rce1 were highly up-regulated at both m RNA and protein level in carcinoma tissues when compared with those of adjacent ones through q RT-PCR and Western blot analysis. By IHC

positive expression of Rce1 was examined in55/76(70.5%)of UCBs and in 4/21(19.0%)of paired pericarcinoma tissues. Positive expression of Rce1 was correlated with poorer differentiation

higher T classification and tumor metastasis(P<0.05). In Kaplan-Meier survival analysis

significant association between positive expression of Rce1 and shortened patients’ survival was found(P<0.001). Compared with those in control group and negative control group

expressions of Rce1 m RNA and protein were significantly reduced after Rce1-sh RNA lentiviral interference vectors being infected(P=0.000)and apoptosis was increased significantly(P=0.000). Conclusion:The expression of Rce1 is significantly higher in UCBs when compared with that of pericarcinoma tissues;positive expression of Rce1 also correlated with prognosis of patients with UCB. Lentivirus-mediated sh RNA silencing Rce1 gene can down regulate the expression of Rce1 gene

promote apoptosis in T24 cells. Rce1 may serve as a biomarker of diagnosis and prognosis in patients with UCB.

关键词

膀胱癌Rce1免疫组化生存分析

Keywords

bladder urothelial carcinomaRce1immunohistochemistrysurvival analysis

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