人源性RUNX相关转录因子1在肾癌中的表达及生物学功能  

Expression and biological function of human-derived RUNX1 gene in renal cancer

作　　者：蔡波[1] 邢钱伟[1] 吴优[1] 管杨波[1] 郭新[1] CAI Bo;XING Qian-wei;WU You;GUAN Yang-bo;GUO Xin(Department of Urology,The Hospital Affiliated to Nantong University,Nantong 226001,Jiangsu,China)

机构地区：[1]南通大学附属医院泌尿外科,南通226001

出　　处：《医学研究生学报》2020年第5期509-514,共6页Journal of Medical Postgraduates

摘　　要：目的人源性RUNX相关转录因子1(RUNX 1)在肾癌中的作用报道较少。文中旨在研究RUNX1基因在肾癌中的表达及生物学功能。方法从GEO数据库下载GSE66272肾癌芯片原始CEL文件,包括肾癌及癌旁组织样本,利用R软件对组织样本基因进行差异表达分析。选取2015年1月至2019年6月南通大学附属医院泌尿外科20份肾癌组织标本,另取对应癌旁组织标本。基因芯片分析RUNX 1在肾癌组织中的表达。小干扰RNA转染786-O细胞株,敲低RUNX1基因,将细胞分为:siRNA1组(转染si-RUNX1的siRNA1序列)、siRNA2组(转染si-RUNX1的siRNA2序列)、siRNA3组(转染si-RUNX1的siRNA3序列)、对照组(对照序列空载体siRNA转染)。实时定量PCR测定RUNX 1含量;细胞克隆形成实验统计细胞克隆形成数量;MTT法检测786-O细胞增殖活性;Transwell肿瘤细胞侵袭实验分析细胞迁移数量;Western blot检测蛋白水平变化。结果RUNX 1在肾癌组织中的表达水平(1.95±0.09)显著高于癌旁组织(0.62±0.05),差异有统计学意义(P<0.01)。siRNA1组、siRNA2组、siRNA3组的siRNA敲减效率显著低于对照组(P<0.01)。MTT检测结果显示,siRNA1组较对照组显著抑制786-O肾癌细胞的增殖(P<0.01)。克隆形成结果显示,siRNA1组786-O肾癌细胞的集落形成效率较对照组显着降低(P<0.003)。siRNA1组的细胞过膜数量[(98.67±3.53)个/视野]明显低于对照组[(143.3±8.74)个/视野],差异有统计学意义(P<0.01)。Western blot结果显示,siRNA1组RUNX1、N-cadherin、MMP9、p-AKT及p-GSK3β较对照组明显下降,E-cadherin明显增加。结论RUNX1在肾癌组织的高表达可促进肾癌细胞的增殖和迁移,故RUNX 1可作为肾癌的潜在临床诊治靶点及预后标志物。Objective It remains an open question of whether the human-derived RUNX-related transcription factor 1(RUNX1)influences the development of renal cell carcinoma.This study aims to investigate the expression and biological function of the RUNX1 gene in renal cell carcinoma.Methods Bioinformatics technique of gene chip was used to identify the expression of RUNX1 in renal cancer.The expression level of RUNX1 in renal cancer tissue was determined by real-time polymerase chain reaction(RT-PCR).Twenty samples of cancer tissue were collected from the Department of Urology,Affiliated Hospital of Nantong University between January 2015 and June 2019.Accordingly,the adjacent normal tissue of the tumor was as well collected.The 786-O cell line was transfected using small interfering RNA,and was subsequently divided into three groups by knocking down the RUNX1 gene:siRNA1 group(siRNA1 sequence transfected with si-RUNX1),siRNA2 group(siRNA2 sequence transfected with si-RUNX1),siRNA3 Group(siRNA3 sequence transfected with si-RUNX1),control group(control sequence empty vector siRNA transfection).Cell clone formation experiment was used to count the number of cell clone formation;MTT assay was used to detect 786-O cell proliferation activity;the Transwell tumor cell invasion experiment was used to analyze the amount of cell migration;Western blot was used to detect changes in protein levels.Results The expression of RUNX1 in renal tumor tissue was significantly higher than that in adjacent normal tissue.The expression of RUNX1 in renal tumor tissues was increased with the escalation of the malignant degree of the pathological stage.The prognosis of patients with high expression of RUNX1 was significantly poor than that of the patients with low expression of RUNX1.The results of cell colony formation assay and MTT assay showed that the cell viability and proliferation of si-RUNX1 groups were significantly inhibited compared to the control group(P<0.01 for both).Transwell assay showed that the number of 786-O cells passing through

关 键 词：人源性RUNX相关转录因子1 肾癌 细胞增殖 细胞迁移 

分 类 号：R737.11[医药卫生—肿瘤]