肾透明细胞癌中miR-200c负性调控ZEB2基因表达的机制研究  

The mechanism of miR- 200c negatively regulating ZEB2 gene in renal clear cell carcinoma

作　　者：司青乐 张辉[1] 郭启振[1] 

机构地区：[1]中国医科大学附属盛京医院泌尿外科,辽宁沈阳110004

出　　处：《现代肿瘤医学》2015年第20期2967-2969,共3页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:30901480);辽宁省科学技术计划项目(编号:2013408001)

摘　　要：目的:探讨肾透明细胞癌中miR-200c调控ZEB2基因表达的分子机制。方法:实时定量PCR法检测肾透明细胞癌及癌旁组织中miR-200c及ZEB2基因的表达水平。培养肾透明细胞癌Caki-1细胞,将miR-200c的前体转染Caki-1细胞,Western blot法检测ZEB2蛋白的表达改变,荧光素酶报告基因表达分析实验验证miR-200c与ZEB2基因的3'非翻译区(3'UTR)的结合及调控作用。结果:实时定量PCR表达检测显示,与癌旁组织相比,miR-200c在肾透明细胞癌组织中的表达显著下调,而ZEB2 mRNA在肾透明细胞癌组织中的表达则呈现明显上调。Pearson相关性分析结果表明,在肾透明细胞癌及癌旁组织中miR-200c与ZEB2基因的表达均显示负性相关。荧光素酶报告基因表达分析实验明确miR-200c能够与ZEB2基因的3'UTR特异性地结合,并抑制荧光素酶的表达。miR-200c前体能够显著下调Caki-1细胞中ZEB2蛋白的表达。结论:miR-200c与ZEB2基因的表达异常与肾透明细胞癌相关,在肾透明细胞癌细胞中miR-200c能够负性调节其靶基因ZEB2的表达。Objective:To explore the molecular mechanism of miR -200c regulating ZEB2 gene in renal clear cell carcinoma.Methods:Real -time PCR was used to detect the expression of miR -200c and ZEB2 mRNA in renal clear cell carcinoma and its adjacent tissues.Western blot assay was used to investigate the ZEB2 protein level and lu-ciferase assay was used to verify the binding ability of miR -200c on ZEB2 after miR -200c precursor transfected in-to Caki -1 cells.Results:Real -time PCR results showed that miR -200c down -regulated in renal clear cell carci-noma tissues compared to its adjacent tissues,and ZEB2 mRNA up -regulated in renal clear cell carcinoma tissues. The Pearson relative analysis showed that miR -200c was negatively related to the ZEB2 gene in renal clear cell car-cinoma and its adjacent tissues.Western blot and luciferase assay revealed that miR -200c could negatively regulate ZEB2 protein expression by binding to the 3'UTR of ZEB2 gene.Conclusion:The abnormal expression of miR -200c and ZEB2 involved in renal clear cell carcinoma and miR -200c could negatively target and silence ZEB2 gene ex-pression in renal clear cell carcinoma cells.

关 键 词：肾透明细胞癌 MIR-200C ZEB2 Caki-1细胞 

分 类 号：R737.11[医药卫生—肿瘤]