LncRNA CASC7通过调控miR-19b-3p/SMG1轴抑制肝癌细胞的增殖、迁移和侵袭  

LncRNA CASC7 inhibits proliferation,migration and invasion of hepatoma cells by regulating miR-19b-3p/SMG1 axis

作　　者：秦宝山[1] 郭云霞[1] 王勇[1] 马英杰[1] QIN Bao-shan;GUO Yun-xia;WANG Yong;MA Ying-jie(Department of Gastroenterology,People’s Hospital of Zhengzhou,Zhengzhou,Henan 450003,China)

机构地区：[1]郑州人民医院消化内科,河南郑州450003

出　　处：《热带医学杂志》2021年第3期276-283,343,共9页Journal of Tropical Medicine

基　　金：河南省医学科技攻关计划项目(2018010044)。

摘　　要：目的探讨长链非编码RNA(lncRNA)癌易感性候选基因7(CASC7)对肝癌细胞增殖、迁移和侵袭的影响及作用机制。方法实时定量聚合酶链式反应(RT-qPCR)检测肝癌组织、对应癌旁组织和肝癌细胞系Huh7、HepG2和MHCC97H及正常肝细胞系THLE-2中CASC7、微小RNA-19b-3p(miR-19b-3p)和生殖器形成抑制基因1(SMG1)mRNA表达水平。以肝癌HepG2细胞为研究对象,转染CASC7过表达载体、miR-19b-3p抑制剂或SMG1过表达载体至HepG2细胞,3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)、Transwell和蛋白印迹(Western blot)分别检测过表达CASC7、沉默miR-19b-3p表达或过表达SMG1对HepG2细胞增殖、迁移和侵袭及细胞周期蛋白D1(Cyclin D1)、细胞周期依赖性蛋白激酶抑制因子1A(p21)、基质金属蛋白酶-2(MMP-2)和E-钙粘附素(Ecadherin)蛋白表达的影响。双荧光素酶报告基因实验验证CASC7与miR-19b-3p及miR-19b-3p与SMG1之间的调控关系。结果与癌旁组织比较,肝癌组织中CASC7和SMG1 mRNA水平降低,miR-19b-3p水平升高,差异均有统计学意义(P<0.05)。与THLE-2细胞比较,肝癌细胞系Huh7、HepG2和MHCC97H中CASC7和SMG1 mRNA水平降低,miR-19b-3p水平升高,差异均有统计学意义(P<0.05)。过表达CASC7、沉默miR-19b-3p表达或过表达SMG1均可降低HepG2细胞存活率、迁移和侵袭数及Cyclin D1和MMP-2蛋白表达,提高p21和E-cadherin蛋白表达,差异均有统计学意义(P<0.05)。CASC7负调控miR-19b-3p表达,miR-19b-3p负调控SMG1表达。过表达miR-19b-3p或沉默SMG1表达逆转了过表达CASC7对HepG2细胞存活率、迁移和侵袭及相关蛋白Cyclin D1、p21、MMP-2和Ecadherin表达的影响。结论过表达CASC7可能通过调控miR-19b-3p/SMG1轴抑制肝癌细胞的增殖、迁移和侵袭。Objective To investigate the effects of IncRNA cancer susceptibility candidate 7(CASC7)on proliferation,migration and invasion of hepatoma cells and its mechanism.Methods RT-qPCR was used to detect the expression levels of CASC7,miR-19 b-3 p and suppressor of morphogenesis of genitalia 1(SMG1)mRNA in hepatocarcinoma tissues,corresponding adjacent tissues,hepatoma cell lines(Huh7,HepG2 and MHCC97 H),and normal liver cell line THLE-2.After the CASC7 overexpression vector,miR-19 b-3 p inhibitor or SMG1 overexpression vector was transfected into HepG2 cells,3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT),Transwell and Western blot were used to detect the effects of over-expressing CASC7,silencing miR-19 b-3 p or over-expressing SMG1 on proliferation,migration and invasion of HepG2 cells and the protein expression levels of Cyclin D1,cyclin-dependent kinase inhibitor 1 A(p21),matrix metalloproteinase-2(M MP-2)and E-cadherin,respectively.The dual luciferase reporter gene assay verified the regulatory relationship between CASC7 and miR-19 b-3 p or miR-19 b-3 p and SMG1.Results Compared with adjacent tissues,the levels of CASC7 and SMG1 mRNA in liver cancer tissues decreased(P<0.05),while the level of miR-19 b-3 p increased(P<0.05).Compared with THLE-2 cells,the levels of CASC7 and SMG1 mRNA in liver cancer cell lines Huh7,HepG2 and MHCC97 H decreased(P<0.05),while the levels of miR-19 b-3 p increased(P<0.05).After over-expressing CASC7,silencing miR-19 b-3 p or over-expressing SMG1,HepG2-cells survival,number of migrated and invaded cells,and the protein expression levels of Cyclin D1 and MMP-2 were reduced(P<0.05),while the protein expression of p21 and Ecadherin increased(P<0.05).CASC7 negatively regulated the expression of miR-19 b-3 p,and miR-19 b-3 p negatively regulated the expression of SMG1.Over-expressing miR-19 b-3 p or silencing SMG1 reversed the effects of over-expressing CASC7 on HepG2 cells survival,number of migrated and invaded cells,and the protein expression of Cyclin D1,p21,MMP-2 and

关 键 词：肝癌 CASC7 miR-19b-3p 生殖器形成抑制基因1 细胞增殖 迁移 侵袭 

分 类 号：R735.7[医药卫生—肿瘤]