miR-593-5p调控肝癌细胞增殖、迁移及侵袭的机制  

Mechanism of miR-593-5p in regulating the proliferation,migration and invasion of hepatoma cells

作　　者：钟腾猛[1] 黄俊玲[3] 李广志[4] 陆炳站 ZHONG Tengmeng;HUANG Junling;LI Guangzhi(Department of Genneral Surgery,People’s Hospital of Baise City,Guangxi Zhuang Nationality Autonomous Region,Baise 533000,China;不详)

机构地区：[1]广西壮族自治区百色市人民医院肝胆外科、疝与腹壁外科,533000 [2]广西壮族自治区百色市人民医院肝胆外科,533000 [3]右江民族医学院附属医院消化内科,广西壮族自治区533000 [4]右江民族医学院附属医院ICU

出　　处：《河北医药》2020年第20期3050-3054,3059,共6页Hebei Medical Journal

摘　　要：目的探讨微小RNA-593-5p(miR-593-5p)对肝癌细胞增殖、迁移及侵袭的影响及作用机制。方法采用qPCR检测正常肝细胞HL7702和肝癌细胞系Hep3B、HepG2、SMMC-7721、Huh7中miR-593-5p和PHF10 mRNA表达。在SMMC-7721细胞中转染miR-593-5p,MTT法检测细胞增殖,Transwell小室法检测细胞迁移和侵袭,Western blot检测细胞周期蛋白D1(CyclinD1)、p21、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、磷脂酰肌醇-3激酶(PI3K)、磷酸化磷脂酰肌醇-3激酶(p-PI3K)、蛋白激酶B(Akt)和磷酸化蛋白激酶B(p-Akt)蛋白表达。starBase软件结合双荧光素酶报告实验分析miR-593-5p和PHF10靶向关系。miR-593-5p和pcDNA-PHF10共转染,观察PHF10过表达对miR-593-5p过表达诱导的SMMC-7721细胞增殖、迁移、侵袭及PI3K/Akt信号通路的影响。结果miR-593-5p在肝癌细胞系Hep3B、HepG2、SMMC-7721、Huh7中表达下调(P<0.05),PHF10 mRNA表达上调(P<0.05)。miR-593-5p过表达明显降低SMMC-7721细胞24 h、48 h、72 h的细胞活性、迁移细胞数、侵袭细胞数及CyclinD1、Vimentin、p-PI3K和p-Akt蛋白表达量(P<0.05),显著提高p21、E-cadherin蛋白水平(P<0.05),对PI3K和Akt蛋白表达无明显影响。miR-593-5p靶向调控PHF10表达。PHF10过表达逆转miR-593-5p过表达对肝癌SMMC-7721细胞增殖、迁移、侵袭、PI3K/Akt信号通路激活的抑制作用。结论miR-593-5p过表达通过靶向PHF10表达及抑制PI3K/Akt信号通路激活而抑制肝癌细胞增殖、迁移及侵袭。Objective To investigate the effects of microRNA-5935p(miR-593-5p)on the proliferation,migration and invasion of hepatoma cells,and to explore its action mechanism.Methods The qPCR.miR-593-5p was used to detect the expression of normal hepatocytes HL7702 and miR-593-5p as well as PF10 in hepatoma cell lines of Hep3B,HepG2,SMMC-7721 and Huh7.miR-593-5p was transfected into SMMCC-7721 cells.MTT assay was used to detect cell proliferation and Transwell chamber assay was used to detect cell migration and invasion.Moreover,Western blot was used to detect the expression levels of CyclinD1,p21,E-cadherin,Vimentin,PI3K,p-PI3K,Akt and p-Akt protein and StarBase software combined with dual luciferase report experiment was used to analyze the targeting relationship between miR-593-5p and PFH10.In addition,miR-593-5p and PFH10 were co-transfected to observe the effects of PHF10 over expression on proliferation,migration,invasion and PI3K/Akt signaling pathway of SMMC-7721 cells induced by the overexpression of miR-593-5p.Results The expression levels of miR-593-5p were down-regulated in hepatoma cell lines of Hep3B,HepG2,SMMC-7721 and Huh7(P<0.05),however,the expression levels of PHF10 were up-regulated(P<0.05).The overexpression of miR-593-5p obviously decreased the activity of SMMC-7721 cells at 24h,48h and 72h,which decreased the migration cell number,invasive cell number and the expression levels of CyclinD1,Vimentin,pPI3K and pAkt in SMMC7721 cells(P<0.05),however,which significantly increased the expression levels of p21 and Ecadherin proteins(P<0.05).Moreover the overexpression of miR-593-5p had no significant effects on the expression levels of PI3K and Akt proteins.In addition the miR-593-5p was used for a targeted regulation of the expression of PHF10.The overexpression of PHF10 reversed the inhibitory effects of miR-593-5p on the proliferation,migration,invasion and activation of PI3K/Akt signaling pathway in hepatoma cells SMMC-7721.Conclusion The overexpression of miR-593-5p can inhibit the proliferation,migrati

关 键 词：miR-593-5p 肝癌 增殖 PHF10 PI3K/AKT信号通路 

分 类 号：R735.7[医药卫生—肿瘤]