敲减BubR1表达对膀胱癌UMUC3细胞生物学行为的影响  

Influence of knocking down BubR1 expression on biochemical behaviors of UMUC3 bladder cancer cells

作　　者：董丰铭 刘屹立[1] DONG Fengming;LIU Yili(Department of Urology,the Fourth Af filiated Hospital of China Medical Universily,Shenyang,Liaoning 110000,China)

机构地区：[1]中国医科大学附属第四医院泌尿外科,辽宁沈阳110000

出　　处：《中华实用诊断与治疗杂志》2024年第7期668-673,共6页Journal of Chinese Practical Diagnosis and Therapy

摘　　要：目的 探讨敲减BubR1表达对膀胱癌UMUC3细胞增殖、迁移及侵袭能力的影响。方法 自TCGA数据库收集19例膀胱癌患者的癌组织标本和癌旁组织标本的BubR1 mRNA表达数据进行比较。自GEO数据库收集165例膀胱癌患者的癌相关基因GSE13507芯片数据及临床数据,比较膀胱癌组织与癌旁组织、表浅性膀胱癌与侵袭性膀胱癌、低级别膀胱癌与高级别膀胱癌、早期膀胱癌与晚期膀胱癌BubR1 mRNA相对表达量;以165例患者癌组织BubR1 mRNA相对表达量均值为界,比较78例高表达(BubR1 mRNA相对表达量≥8.68)与87例低表达(BubR1 mRNA相对表达量<8.68)者的总体生存率和特异性存活率。将用慢病毒感染的方法构建的稳转UMUC3细胞分为空白组(转染shRNA-NC)、转染1组(转染shRNA-BubR1-1)、转染2组(转染shRNA-BubR1-2),转染48 h,采用实时荧光定量PCR法检测BubR1 mRNA相对表达量,采用Western blot法检测BubR1蛋白相对表达量,选择敲减效率高的转染2组细胞进行后续实验。取空白组和转染2组细胞,采用细胞计数法检测转染后培养第1、2、3、4、5天增殖细胞数,采用细胞划痕实验检测转染后培养12、24 h时细胞划痕宽度,采用Transwell小室实验检测细胞侵袭能力。结果 (1)19例膀胱癌患者癌组织BubR1 mRNA相对表达量(7.00±0.76)高于癌旁组织(1.22±0.38)(t=6.803,P<0.001)。(2)165例膀胱癌患者癌组织BubR1 mRNA相对表达量(8.68±0.08)高于癌旁组织(7.83±0.09)(t=5.862,P<0.001),侵袭性膀胱癌(9.60±0.14)高于表浅性膀胱癌(8.43±0.09)(t=4.207,P<0.001)、高级别膀胱癌(9.27±0.12)高于低级别膀胱癌(8.33±0.09)(t=6.472,P<0.001)、晚期膀胱癌(9.07±0.14)高于早期膀胱癌(8.45±0.09)(t=3.962,P<0.001)。BubR1高表达者总体生存率(42.21%)、膀胱癌特异性存活率(70.50%)均低于低表达者(56.78%、83.26%)(P<0.05)。(3)转染1组、转染2组BubR1 mRNA(0.47±0.02、0.19±0.01)及蛋白(0.42±0.04、0.13±0.02)相对表达量均低于空白�Objective To investigate the influence of knocking down the expression of BubR1 on the proliferation, migration and invasion of UMUC3 bladder cancer cells. Methods The data of BubR1 mRNA expression in bladder cancer tissue samples and paracancerous tissue samples of 19 patients from TCGA database were collected for comparison. The bladder cancer related gene GSE13507 chip data and clinical data of 165 bladder cancer patients from GEO database were collected. The relative expressions of BubR1 mRNA were compared between bladder cancer tissues and paracancerous tissues, between superficial bladder cancer and invasive bladder cancer, between low-grade bladder cancer and high-grade bladder cancer, and between early bladder cancer and advanced bladder cancer. Taking the relative expression of BubR1mRNA of 165 bladder cancer patients as the boundary value, the overall survival rate and bladder cancer specific survival rate were compared between 78patients with highly expressed BubR1 mRNA (≥8.68)and 87patients with lowly expressed BubR1mRNA(<8.68).The stable transfection cell line UMUC3constructed by lentivirus infection method were divided into blank group(transfected with shRNA-NC),transfection group 1(transfected with shRNA BubR1-1),and transfection group 2(transfected with shRNA BubR1-2).After 48-h transfection,the relative expression of BubR1mRNA was detected by real-time fluorescence quantitative PCR,and the relative expression of BubR1protein was detected by Western blot.The cells in transfection group 2with the highest knocking down efficiency were used for subsequent experiments.The number of proliferated cells in blank group and transfection group 2 were detected with cytometry by day 1,2,3,4and 5after transfection,the scratch width after 12and 24hculture was detected by scratch assay,and the cell invasion ability was detected by Transwell chamber assay.Results(1)In 19patients with bladder cancer,the relative expression of BubR1mRNA was higher in the bladder cancer tissues(7.00±0.76)than that in the paracanc

关 键 词：膀胱癌 UMUC3细胞 BUBR1 增殖 迁移 侵袭 

分 类 号：R737.14[医药卫生—肿瘤]