衣被蛋白复合物亚基β-2在膀胱癌中的分子机制研究
Molecular mechanisms of coatomer protein complex subunit β-2 in bladder cancer
石泽玢 1李寿春 1左立 1张力峰2
作者信息
- 1. 南京医科大学附属常州第二人民医院 泌尿外科,江苏 常州 213000
- 2. 南京医科大学附属常州第二人民医院 泌尿外科,江苏 常州 213000;南京医科大学,常州医学中心,江苏 常州 213000
- 折叠
摘要
目的 研究衣被蛋白复合物亚基β-2(coatomer protein complex subunit β2,COPB2)的表达对膀胱癌发病的影响,探讨COPB2在膀胱癌中的分子作用机制.方法 从癌症基因组图谱(the cancer genome atlas,TCGA)数据库下载膀胱癌中COPB2的相关数据,探究COPB2的表达水平与预后关系.根据膀胱癌不同分期将样本进行分组.使用Kaplan-Meier法对不同组别间的生存情况进行分析.使用COPB2-shRNA慢病毒和阴性对照慢病毒感染5637细胞(人膀胱癌细胞),RT-qPCR和蛋白印迹检测细胞中COPB2表达变化.使用平板克隆实验评估细胞的增殖能力.对细胞中p27 kip1、半胱天冬酶-3和多腺苷二磷酸多聚酶的表达水平进行蛋白印迹检测.检测细胞中与上皮-间充质转化(epithelial-to-mesenchymal transition,EMT)相关的N-钙黏蛋白和Snail的表达水平.结果 肿瘤免疫浸润评分数据库(tumor immune estimation resource,TIMER)显示,相比于正常膀胱,COPB2在膀胱癌中过表达.COPB2表达水平越高,患者预后越差(P＜0.05).COPB2敲低后膀胱癌细胞增殖能力减弱,凋亡相关蛋白表达增加,EMT相关蛋白表达减弱.结论 COPB2在膀胱癌中的高表达显著促进膀胱癌发生发展,是一种有前途的膀胱癌治疗靶点.
Abstract
Objective To investigate the impact of coatomer protein complex subunit β2(COPB2)expression on the incidence of bladder cancer and explore the molecular mechanisms of COPB2 in bladder cancer.Methods Relevant data on COPB2 in bladder cancer were downloaded from The Cancer Genome Atlas(TCGA)database to explore the relationship between COPB2 expression levels and prognosis.Kaplan-Meier analysis was performed to assess survival differences among different groups.5637 human bladder cancer cells were infected with COPB2-shRNA lentivirus and a negative control lentivirus.RT-qPCR and Western blot were used to detect changes in COPB2 expression in cells.Cell proliferation capacity was evaluated through a colony formation assay.Protein expression levels of p27 kipl,Cleaved Caspase-3,and Cleaved PARP in cells were determined by Western blot.The expression levels of N-Cadherin and Snail,which are related to epithelial-to-mesenchymal transition(EMT),were also simultaneously assessed in cells.Results The TIMER database showed that COPB2 was overexpressed in bladder cancer compared to normal bladder tissue.Higher COPB2 expression levels were associated with poorer patient prognosis(P＜0.05).Knockdown of COPB2 in bladder cancer cells resulted in reduced cell proliferation,increased expression of apoptosis-related proteins,and decreased expression of EMT-related proteins.Conclusions COPB2 is highly expressed in bladder cancer and significantly promotes the development of bladder cancer.It represents a promising therapeutic target for bladder cancer.
关键词
衣被蛋白复合物亚基β-2/膀胱癌/发病机制/生物标志物/治疗靶点Key words
Coatomer protein complex subunit β2/Bladder cancer/Proliferation/Prognostic biomarkers/Therapeutic target引用本文复制引用
基金项目
国家自然科学基金青年科学项目(81902565)
出版年
2024
NETL
NSTL
万方数据