MiR-122通过靶向SIRT1抑制肝癌细胞的增殖、迁移和侵袭  

MiR-122 inhibits the proliferation,migration and invasion of liver cancer cells by targeting SIRT1

作　　者：马珊 刘妍 李冬娟 MA Shan;LIU Yan;LI Dongjuan(Department of Oncology,Zaozhuang Central Hospital of Shandong Guoxin Yiyang group,277000,China)

机构地区：[1]山东国欣颐养集团枣庄中心医院肿瘤科,277000 [2]山东国欣颐养集团枣庄中心医院老年病科,277000

出　　处：《传染病信息》2023年第4期311-319,共9页Infectious Disease Information

摘　　要：目的探讨微小RNA-122(micro RNA-122,miR-122)通过靶向沉默信息调节因子1(silent information regulator1,SIRT1)对肝癌细胞增殖、迁移和侵袭的影响。方法用实时荧光定量PCR(real-time quantitative PCR,RT-qPCR)法检测肝癌组织和细胞中miR-122和SIRT1 mRNA表达水平;用生物信息学和双荧光素酶报告基因分析并验证miR-122和SIRT1靶向关系。培养肝癌细胞Hep G2,将其分为空白对照组(control组)、mi R-122过表达阴性对照组(miR-NC mimic组)、mi R-122过表达组(miR-122 mimic组)、miR-122过表达和SIRT1过表达阴性对照组(miR-122 mimic+pcDNA组)、mi R-122过表达和SIRT1过表达组(miR-122 mimic+pcDNA-SIRT1组)。用细胞计数试剂(cell counting kit-8,CCK-8)和5-乙炔基-2’脱氧尿嘧啶核苷分析各组细胞增殖情况;划痕愈合实验和Transwell实验分析各组细胞迁移和侵袭能力;用Western blot法分析各组细胞中增殖、迁移和侵袭相关蛋白p53、细胞周期蛋白D1、E-钙粘蛋白、N-钙粘蛋白和波形蛋白及SIRT1蛋白表达水平。结果MiR-122表达水平在肝癌组织和细胞中下调,SIRT1 mRNA表达水平在肝癌组织和细胞中上调(P<0.05)。MiR-122负靶向调控SIRT1表达(P<0.05)。过表达miR-122抑制Hep G2细胞增殖、迁移和侵袭,降低SIRT1、细胞周期蛋白D1、N-钙粘蛋白和波形蛋白蛋白水平,升高p53和E-钙粘蛋白蛋白水平(P<0.05)。然而,SIRT1过表达可部分逆转过表达miR-122对Hep G2细胞的作用(P<0.05)。结论MiR-122通过负靶向调控SIRT1抑制肝癌细胞增殖、迁移和侵袭。Objective To explore the effects of microRNA-122(miR-122)on the proliferation,migration and invasion of liver cancer cells by targeting silent information regulator 1(SIRT1).Methods Real-time quantitative PCR(RT-qPCR)was used to detect the expression levels of miR-122 and SIRT1 mRNA in liver cancer tissues and cells;bioinformatics and dual luciferase reporter gene were used to analyze and verify the targeting relationship between miR-122 and SIRT1.The liver cancer cells HepG2 were cultured and divided into blank control group(control group),miR-122 overexpression negative control group(miR-NC mimic group),miR-122 overexpression group(miR-122 mimic group),miR-122 overexpression and SIRT1 overexpression negative control group(miR-122 mimic+pcDNA group),and miR-122 overexpression and SIRT1 overexpression group(miR-122 mimic+pcDNA-SIRT1 group).Cell counting kit-8(CCK-8)and 5-Ethynyl-2’-deoxyuridine(EdU)methods were used to analyze cell proliferation in each group;scratch healing test and Transwell test were used to analyze cell migration and invasion abilities of each group;Western blot was used to analyze the expression levels of proliferation,migration and invasion related proteins of cells in each group,including p53,CyclinD1,E-cadherin,N-cadherin,vimentin and SIRT1 proteins.Results The expression level of miR-122 was down-regulated in liver cancer tissues and cells,and the expression level of SIRT1 mRNA was up-regulated in liver cancer tissues and cells(P<0.05).MiR-122 negatively targeted the expression of SIRT1(P<0.05).Overexpression of miR-122 inhibited HepG2 cell proliferation,migration and invasion,decreased SIRT1,CyclinD1,N-cadherin and vimentin protein levels,and increased p53 and E-cadherin protein levels(P<0.05).However,overexpression of SIRT1 could partially reverse the effect of overexpression of miR-122 on HepG2 cells(P<0.05).Conclusion MiR-122 inhibits the proliferation,migration and invasion of liver cancer cells by negatively targetingly regulating SIRT1.

关 键 词：微小RNA-122 沉默信息调节因子1 肝癌 HepG2 增殖 迁移 侵袭 细胞周期蛋白D1 E-钙粘蛋白 

分 类 号：R735.7[医药卫生—肿瘤]