【摘要】 目的:探讨冷诱导RNA结合蛋白(cold inducible RNA binding protein,CIRBP)基因在肾癌中的生物学功能。方法:用GEO(Gene Expression Omnibus)和癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库的基因芯片和基因测序数据分析肾癌中CIRBP mRNA的表达水平。实时荧光定量PCR法验证CIRBP基因在20例肾癌组织及3种肾癌细胞株中的表达水平。将CIRBP过表达质粒分别转入至肾癌786-O和CAKI-1细胞,随后采用MTT法和细胞克隆形成实验检测肾癌细胞的增殖活性,并采用Transwell小室实验检测肾癌细胞的迁移能力,最后采用蛋白质印迹法检测细胞迁移相关蛋白N-钙黏蛋白(N-cadherin)和E-钙黏蛋白(E-cadherin)以及蛋白激酶B(protein kinase B,PKB,又称AKT)通路相关蛋白的表达水平。结果:肾癌组织中CIRBP mRNA的表达水平明显低于癌旁正常组织。高表达CIRBP肾癌患者的预后明显优于CIRBP低表达者。CIRBP过表达质粒转染后,肾癌786-O和CAKI-1细胞的增殖活性和克隆形成能力明显受到抑制(P值均<0.01),穿过Transwell小室膜的细胞数明显少于对照组(P值均<0.01)。而且CIRBP基因过表达后,肾癌细胞中迁移相关蛋白N-cadherin表达明显降低,E-cadherin水平明显增高,而AKT通路关键蛋白磷酸化AKT(phospho-AKT,p-AKT)和磷酸化糖原合酶激酶3β(phospho-glycogen synthasc kinase 3β,p-GSK3β)的表达水平明显下调(P值均<0.05)。结论:CIRBP在肾癌组织中表达下调,CIRBP过表达可抑制肾癌细胞的增殖和迁移,提示其可以作为一种肾癌的潜在临床诊治靶点及预后标志物。更多还原

【Abstract】 Objective:To investigate the expression and biological functions of cold inducible RNA binding protein (CIRBP) in renal cancer.Methods:Bioinformatics analysis of microarray in Gene Expression Omnibus (GEO) and gene sequencing data in The Cancer Genome Atlas (TCGA) was used to analyze the expression of CIRBP mRNA in renal cancer,further the expression level of CIRBP gene in 20 cases of renal cancer tissues and 3 kinds of renal cancer cell lines was identified by real-time fluorescent quantitative PCR.The renal cancer 786-O and CAKI-1 cells were transfected with the CIRBP overexpression plasmids,then the cell proliferation viability was detected by MTT assay and cell clone formation assay.The migration ability of renal cancer cells was detected by Transwell chamer,and the expressions of cell migration-related protein N-cadherin,E-cadherin and protein kinase B (PKB,also known as AKT) pathway-related proteins were detected by Western blotting.Results:The expression level of CIRBP in renal cancer tissues was significantly lower than that in the adjacent normal tissues.The prognosis of patients with high expression of CIRBP mRNA was significantly better than that of the patients with low expression of CIRBP.The proliferation and clone formation of renal cancer 786-O and CAKI-1 cells transfected with CIRBP overexpresion plasmids were significantly inhibited (all P < 0.01).The number of renal cancer 786-O and CAKI-1 cells migrated through the membrane in CIRBP overexpression group was less than that in the control group (all P < 0.01).In the 786-O and CAKI-1 cells with CIRBP overexpression,the expression level of migration-related protein N-cadherin was significantly decreased,the expession level of E-cadherin was significantly increased,while the expressions of AKT pathway-related phospho-AKT (p-AKT) and phospho-glycogen synthasc kinase 3β (p-GSK3β) proteins were decreased significantly (all P < 0.05).Conclusion:CIRBP is down-regulated in renal cancer,and inhibits the proliferation and migration of renal cancer cells.CIRBP can be used as a potential clinical diagnosis target and prognostic marker for renal cancer.更多还原