miR-450b-5p在肝细胞癌中的表达及功能  

The expression and function of miR-450b-5p in hepatocellular carcinoma

作　　者：张雷[1] 王宇锋[2] 王亮[2] 殷国志[2] 韩少山[2] 李瑞祥[1] 鲜瑶[3] 姚德茂[1] ZHANG Lei;WANG Yu-feng;WANG Liang;YIN Guo-zhi;HAN Shao-shan;LI Rui-xiang;XIAN Yao;YAO De-mao(Department of Geriatric Surgery, 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;Department of Nutrition, 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老年外科,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 [3]西安交通大学第一附属医院营养科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2019年第5期747-754,共8页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：陕西省自然科学基础研究计划(No.2017JM8002,No.2016JQ8029)~~

摘　　要：目的探究miR-450b-5p在肝细胞癌(hepatocellular carcinoma,HCC)中的表达情况、临床意义及生物学功能,并初步探究其潜在的作用机制。方法Real-time PCR检测miR-450b-5p在HCC组织和细胞系的表达情况;分析miR-450b-5p的表达水平与HCC临床病理特征及患者预后的相关性,并用Cox多因素回归分析法分析患者的独立预后因素;采用Transwell迁移和侵袭实验检测miR-450b-5p对HCC细胞迁移与侵袭能力的影响;MTT实验观察miR-450b-5p对HCC细胞增殖能力的影响;应用生物信息学工具预测miR-450b-5p的下游靶点,并应用荧光素酶报告实验及Western blot分析miR-450b-5p对靶点的调节作用。结果 miR-450b-5p在HCC组织和细胞系中的表达均明显降低(均P<0.001);miR-450b-5p的表达水平与肿瘤大小(P=0.026)、静脉侵犯(P=0.013)、TNM分期(P=0.020)及HCC患者的预后(P=0.014 2)相关,且Cox多因素回归分析结果显示miR-450b-5p为HCC患者的独立预后因素之一;上调MHCC-97H细胞的miR-450b-5p表达后,细胞迁移、侵袭和增殖能力显著被抑制;而下调Hep3B的miR-450b-5p表达时,细胞的迁移、侵袭和增殖能力明显增强;生物信息学分析发现,性别决定区Y框蛋白2(sex determining region Y-box2,SOX2)可能是miR-450b-5p的作用靶点;荧光素酶报告实验显示,miR-450b-5p能够负向调节SOX2-3′UTR的荧光素酶活性;miR-450b-5p mimics能降低MHCC-97H细胞中SOX2的表达水平,miR-450b-5p inhibitors能上调Hep3B细胞中SOX2的表达水平。结论 miR-450b-5p通过靶向作用于SOX2抑制HCC细胞的迁移、侵袭及增殖,进而抑制HCC的发生发展。Objective To investigate the expression, clinical significance and biological function of miR-450b-5p in hepatocellular carcinoma (HCC) so as to make a preliminary exploration of the mechanism in HCC.Methods Real-time PCR was used to detect miR-450b-5p expression in HCC tissues and cells.Statistical analysis was employed to explore the correlations of miR-450b-5p with clinicopathologic features and prognosis.And Cox regression analysis was applied to analyze the relationship between clinicopathologic characteristics and prognosis.Transwell assay and MTT assay were used to study the effects of miR-450b-5p on migration, invasion and proliferation of HCC cells.Bioinformatics tools were applied to predict the potential target gene of miR-450b-5p.Luciferase reporter assay and Western blot were employed to determine the correlation between miR-450b-5p and its target gene.Results miR-450b-5p was significantly down-regulated in HCC tissues and cell lines ( P <0.001).miR-450b-5p expression was significantly correlated with tumor size ( P =0.026), portal vein infiltration ( P = 0.013 ), TNM stage ( P =0.020), and the prognosis of HCC patients.Cox regression analysis showed that miR-513a-5p expression was an independent prognostic factor for the HCC patients survival.Overexpressed miR-450b-5p notably inhibited migration, invasion and proliferation of MHCC-97H cells, while miR-450b-5p inhibitors had the opposite effects on Hep3B cells.Bioinformatics analysis suggested that Sex Determining Region Y Box 2 (SOX2) might be the target of miR-450b-5p.Furthermore, luciferase reporter assay indicated that the luciferase activity of SOX2-3’-UTR was negatively regulated by miR-450b-5p.Consistently, Western blot showed that miR-450b-5p negatively regulated the expression of SOX2 in HCC cells.Conclusion miR-450b-5p suppresses migration, invasion and proliferation of HCC cells by targeting SOX2.

关 键 词：肝细胞癌 miR-450b-5p SOX2 迁移 侵袭 增殖 

分 类 号：R735.7[医药卫生—肿瘤]