LRRC59促进膀胱尿路上皮癌细胞增殖、侵袭及凋亡抑制  

LRRC59 promotes proliferation,invasion and inhibition of apoptosis in bladder urothelial carcinoma cells

作　　者：张鹏[1] 梁培禾[1] ZHAGN Peng;LIANG Peihe(Department of Urology,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400010,China)

机构地区：[1]重庆医科大学附属第二医院泌尿外科,重庆400010

出　　处：《陆军军医大学学报》2023年第14期1526-1538,共13页Journal of Army Medical University

基　　金：重庆市中青年医学高端后备人才项目(第四批);重庆市基础科学与前沿技术专项面上项目(CSTC2017jcyjAX0435);重庆市留学人员回国创业创新支持计划(CX2019146)。

摘　　要：目的探讨富含亮氨酸重复序列蛋白59(leucine-rich repeat-containing protein 59,LRRC59)在膀胱尿路上皮癌中的表达及其对肿瘤细胞增殖、凋亡及侵袭能力的影响。方法收集重庆医科大学附属第二医院泌尿外科2020年10月至2021年9月膀胱尿路上皮癌患者组织标本10例,使用免疫组化技术分析膀胱癌组织及癌旁组织中LRRC59表达情况。用在线数据库(TIMER2.0、GEPIA2、Linked Omics)分析泛癌、膀胱癌及正常膀胱中LRRC59的表达情况并分析其预后,通过分析膀胱癌中具有关联性的基因进行通路富集分析。RT-qPCR及Western blot检测膀胱尿路上皮癌细胞系T24、5637、J82和正常尿路上皮细胞系SV-HUC-1中LRRC59表达,以SV-HUC-1细胞作为对照。选择T24及5637细胞系,通过质粒转染shRNA下调LRRC59表达,以shRNA-NC作为对照组,shRNA-1、shRNA-2、shRNA-3作为实验组。CCK8和克隆形成实验检测细胞增殖能力,Transwell和细胞划痕实验检测细胞迁移、侵袭能力,AnnexinV-FITC、碘化丙啶(PI)染色后流式细胞术检测细胞凋亡和周期,Western blot检测上皮间质转化、凋亡及p53通路相关蛋白的表达。结果免疫组化结果显示LRRC59在膀胱尿路上皮癌组织中的表达高于癌旁组织(P<0.05);数据库检索显示LRRC59在膀胱癌患者中表达高于正常者(P<0.05),且患者中高表达者预后更差(P<0.01)。通路富集分析提示LRRC59相关基因主要富集在内质网蛋白质加工、细胞周期与DNA复制;敲低LRRC59的表达能抑制膀胱癌细胞的增殖,细胞发生G1期阻滞,凋亡率增加,促凋亡蛋白Bax表达增高,抑凋亡蛋白Bcl-2表达下降;细胞迁移及侵袭能力下降。上皮-间质转化相关分子检测提示Snail、Vimentin蛋白表达下降,而E-cadherin蛋白表达增高(P<0.05)。p53通路蛋白检测提示p53、p21蛋白表达升高,而Cyclin D1蛋白表达下降(P<0.05)。结论LRRC59的表达促进膀胱尿路上皮癌细胞增殖、迁移和侵袭;LRRC59可能是膀�Objective To investigate the expression of leucine-rich repeat-containing protein 59(LRRC59)in bladder urothelial carcinoma and its effect on proliferation,apoptosis and invasion of tumor cells.Methods Ten tissue samples were collected from the patients with bladder urothelial carcinoma hospitalized in our hospital from Oct.2020 to Sep.2021,and then the expression of LRRC59 in the tumor and adjacent non-cancerous tissues was measured with immunohistochemical assay.The expression of LRRC59 in pan-carcinoma,bladder cancer and normal bladder was also analyzed in Online databases TIMER,GEPIA and LinkedOmics,and its relationship with prognosis of these subjected patients was analyzed.Then pathway enrichment analysis was performed by analyzing the genes related to bladder cancer.LRRC59 expression in bladder urothelial carcinoma cell lines T24,5637 and J82 and human bladder epithelial cell line SV-HUC-1(as control)was detected by RT-qPCR and Western blotting.After the expression of LRRC59 in T24 and 5637 cells were knocked down by sh-RNA techniques(shRNA-1,shRNA-2 and shRNA-3 for knockdown,and shRNA-NC as blank control).Then cell proliferation was detected by CCK8 assay and colony formation assay,migration and invasion were observed by Transwell assay and cell scratch assay,and cell apoptosis and cycle were measured by flow cytometry after AnnexinV-FITC and Propidium Iodide(PI)staining.The expression of epithelial-mesenchymal transition(EMT)related proteins,apoptosis-related proteins and p53 pathway-related proteins were detected by Western blotting.Results The expression of LRRC59 was higher in bladder urothelial carcinoma tissues than adjacent non-cancerous tissues(P<0.05).Database analysis showed that LRRC59 expression was higher in the bladder cancer tissue than the normal tissues(P<0.01),and the patients with high LRRC59 expression had a worse prognosis(P<0.01).Pathway enrichment analysis suggested that LRRC59-related genes were mainly enriched in protein processing in endoplasmic reticulum,cell cycle and DNA repli

关 键 词：膀胱癌 LRRC59 增殖 凋亡 上皮间质转化 

分 类 号：R341[医药卫生—基础医学] R730.23R737.14