miR-3127-5p靶向抑制Cdk10表达促进肝癌细胞迁移与侵袭  

miR-3127-5p promotes hepatocellular carcinoma cell migration and invasion through targeting Cdk10

作　　者：丁琭 王亮[2] 石磊[2] DING Lu;WANG Liang;SHI Lei(Department of Anesthesiology,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China;Department of Hepatobiliary Surgery,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China)

机构地区：[1]西安交通大学第一附属医院麻醉科,陕西西安710061 [2]西安交通大学第一附属医院肝胆外科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2020年第1期90-96,共7页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省重点研发计划(No.2015SF039)~~

摘　　要：目的探究miR-3127-5p在肝细胞癌(hepatocellular carcinoma,HCC)中的表达情况、临床意义及作用机制。方法实时定量PCR(qPCR)检测miR-3127-5p在HCC组织与癌旁组织、正常肝细胞(L02)与HCC细胞系中的表达情况;TCGA数据库分析正常肝组织及肝癌组织中miR-3127-5p的表达差异;分析miR-3127-5p的表达与HCC患者临床病理特征及预后的关系;在HCCLM3细胞中敲减miR-3127-5p,Transwell实验来检测细胞侵袭及迁移能力;TargetScan数据库预测miR-3127-5p下游靶基因,并通过双荧光素酶报告基因实验加以验证。结果与癌旁组织和正常肝细胞相比,miR-3127-5p在HCC组织及细胞系中均高表达(P<0.05),与门静脉癌栓(P=0.016)及TNM分期(P=0.016)有关;基于TCGA数据库数据的生存分析显示,miR-3127-5p高表达与不良预后相关(P=0.002);在HCCLM3细胞中敲减miR-3127-5p抑制细胞的侵袭及迁移;通过生物信息学预测并通过双荧光素酶报告基因实验证实细胞周期素依赖性激酶10(cyclin-dependent kinase 10,Cdk10)是miR-3127-5p的直接靶基因,回复实验证实Cdk10介导了miR-3127-5p对肝癌细胞侵袭及迁移能力的促进作用。结论在HCC中miR-3127-5p通过靶向抑制抑癌基因Cdk10从而促进肝癌细胞侵袭及迁移。Objective To investigate the expression level,clinical significance and mechanisms of miR-3127-5p in hepatocellular carcinoma(HCC).Methods Real-time quantitative PCR(qPCR)was performed to detect miR-3127-5p expression in HCC tissues and the adjacent non-tumor tissues as well as in HCC cell lines and normal hepatic cells.The difference in miR-3127-5p expresssion between normal liver tissues and HCC tissues was analyzed by TCGA database.The relationship of miR-3127-5p expression with the clinicopathologic features and prognosis of HCC patients was analyzed.After the HCCLM3 cells were transfected with miR-3127-5p inhibitors or negative control sequence,the abilities of cell migration and invasion were measured by Transwell assay;the potential downstream of miR-3127-5p was predicted by TargetScan and validated by Luciferase reporter assay.Results Compared with that in tumor adjacent tissues or normal hepatic cells,miR-3127-5p was increased in HCC tissue and cell lines(all P<0.05).The expression of miR-3127-5p was significantly related to the portal vein tumor thrombus(P=0.016)and advanced TNM stage(P=0.016).The result of overall survival investigated by TCGA database showed that the survival rate in patients with high miR-3127-5p expression was significantly lower than that in those with low miR-3127-5p expression(P=0.0023).Knockdown of miR-3127-5p significantly inhibited HCCLM3 cell migration and invasion compared with negative controls.Cyclin-dependent kinase 10(Cdk10),the potential downstream of miR-3127-5p,was validated by luciferase reporter assay.Rescue assay showed that Cdk10 mediated the effects of miR-3127-5p on migration and invasion of HCC cells.Conclusion miR-3127-5p is upregulated in HCC,which promotes HCC cell migration and invasion through targeting Cdk10.

关 键 词：miR-3127-5p 肝细胞癌 Cdk10 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]