miR-146b-3p通过靶向BTG2基因调控肝癌细胞增殖、侵袭和迁移的机制研究  

Mechanism of miR-146b-3p regulating proliferation,invasion and migration of hepatoma cells by targeting BTG2 gene

作　　者：贺丽娜[1] 刘力[1] 贺晓旭 王建钧[1] 何松[1] HE Li-na;LIU Li;HE Xiao-xu(Department of Hepatobiliary Surgery,Affiliated Hospital of Xiangnan University(Chenzhou,423000),China)

机构地区：[1]湘南学院附属医院肝胆外科,湖南郴州423000

出　　处：《中西医结合肝病杂志》2021年第9期816-821,共6页Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases

摘　　要：目的:探讨miR-146b-3p对肝癌细胞增殖、侵袭和迁移的影响及其作用机制。方法:采用实时荧光定量PCR(qPCR)检测人正常肝细胞系L025和人不同肝癌细胞系Huh7、MHCC97-H、HCCLM3中miR-146b-3p的表达水平,免疫印迹法(Western blot)检测B细胞易位基因2(BTG2)蛋白表达水平。在肝癌Huh7细胞中转染miR-146b-3p inhibitor干扰miR-146b-3p的表达,通过噻唑蓝(MTT)和Transwell实验分析干扰miR-146b-3p对Huh7细胞增殖、侵袭和迁移的影响。生物信息学和双荧光素酶报告基因实验检测miR-146b-3p和BTG2的靶向关系。功能恢复实验检测沉默BTG2对miR-146b-3p调控的Huh7细胞增殖、侵袭和迁移的影响。结果:与人正常肝细胞系比较,人肝癌细胞系Huh7、MHCC97-H、HCCLM3中miR-146b-3p的表达水平明显上调(P<0.05),BTG2的表达明显下调(P<0.05)。干扰miR-146b-3p能够上调BTG2的表达。生物信息学预测显示BTG2可能是miR-146b-3p的直接靶基因,双荧光素酶报告基因实验证实了二者的靶向关系。干扰miR-146b-3p能够抑制Huh7细胞增殖、侵袭和迁移。沉默BTG2能够逆转干扰miR-146b-3p导致的Huh7细胞增殖、侵袭和迁移的抑制作用。结论:干扰miR-146b-3p通过靶向上调BTG2的表达抑制肝癌Huh7细胞增殖、侵袭和迁移。Objective:To investigate the effects of miR-146b-3p on proliferation,invasion,and migration of hepatoma cells and its mechanism.Methods:Real-time quantitative PCR(qPCR)was used to detect the expression of miR-146b-3p in human normal liver cell line L025 and human hepatoma cell lines Huh7,MHCC97-H,and HCCLM3.Western blot was used to detect B cells.Gene 2(BTG2)protein expression level.Transfection of miR-146b-3p inhibitors in hepatocellular carcinoma Huh7 cells interfered with the expression of miR-146b-3p and interfered with the effect of miR-146b-3p on proliferation,invasion,and migration of Huh7 cells by thiazolyl blue(MTT)and Transwell assay.Bioinformatics and dual luciferase reporter assays detect the targeting relationship of miR-146b-3p and BTG2.Functional recovery assay was performed to detect the effect of silencing BTG2 on proliferation,invasion,and migration of HuR7 cells regulated by miR-146b-3p.Results:Compared with human normal liver cell lines,the expression levels of miR-146b-3p in human hepatoma cell lines Huh7,MHCC97-H,and HCCLM3 were significantly up-regulated(P<0.05),and the expression of BTG2 was down-regulated(P<0.05).Interference with miR-146b-3p was able to up-regulate the expression of BTG2.Bioinformatics predictions suggest that BTG2 may be a direct target gene of miR-146b-3p,and dual luciferase reporter gene experiments confirmed the targeting relationship between the two.Interference with miR-146b-3p inhibited Huh7 cell proliferation,invasion,and migration.Silencing BTG2 reversed the inhibition of proliferation,invasion,and migration of Huh7 cells induced by miR-146b-3p.Conclusion:Interfering with miR-146b-3p inhibits the proliferation,invasion,and migration of hepatoma Huh7 cells by targeting up-regulation of BTG2 expression.

关 键 词：肝癌细胞 miR-146b-3p BTG2基因 增殖 侵袭 迁移 

分 类 号：R575.2[医药卫生—消化系统]