LncRNA SNHG6调控miR-186表达对肝癌细胞增殖、迁移和侵袭的影响及机制  

The effect and mechanism of LncRNA SNHG6 regulating miR-186 expression on the proliferation,migration and invasion of hepatocellular carcinoma cells

作　　者：杨鹏生 宋黎明 段希斌[1] 梁马可 梁占强 李学民[1] Yang Pengsheng;Song Liming;Duan Xibin;Liang Make;Liang Zhanqiang;Li Xuemin(Hepatobiliary and Pancreatic Surgery,Zhengzhou Central Hospital,Henan Zhengzhou 450000,China)

机构地区：[1]郑州市中心医院肝胆胰外科,河南郑州450000

出　　处：《现代肿瘤医学》2020年第19期3319-3324,共6页Journal of Modern Oncology

基　　金：河南省科技厅科技攻关项目(编号:182102310482)。

摘　　要：目的:研究SNHG6通过调控miR-186表达对人肝癌细胞增殖、迁移、侵袭和上皮间质转化(EMT)的影响。方法:通过实时荧光定量PCR(RT-qPCR)检测57名肝癌患者癌组织和肝癌细胞系中SNHG6和miR-186的表达;双荧光素酶报告实验验证SNHG6和miR-186的靶向调控关系;将SNHG6的小干扰RNA(si-SNHG6组)、阴性无意义对照序列(si-con组)、si-SNHG6与anti-miR-186抑制剂(si-SNHG6+anti-miR-186组)、si-SNHG6与miR-186抑制物阴性对照(si-SNHG6+anti-miR-con组),均以脂质体法转染至肝癌HepG2细胞,MTT法检测细胞增殖;Transwell检测肝癌细胞迁移和侵袭;Western blot实验检测肝癌细胞CDK4、Cyclin D1、MMP-2、MMP-9及EMT的标志物(E-cadherin、N-cadherin和Vimentin)表达。结果:与癌旁正常组织或正常肝细胞相比,SNHG6在肝癌患者和肝癌细胞系中的表达升高,而miR-186表达降低,两者存在负相关性。与si-con组比较,si-SNHG6组HepG2细胞活力明显下降,迁移和侵袭细胞数明显减少,CDK4、Cyclin D1、MMP-2、MMP-9、N-cadherin和Vimentin蛋白表达降低,E-cadherin蛋白表达增加。SNHG6靶向负调控miR-186表达,抑制miR-186表达可部分逆转下调SNHG6对HepG2细胞增殖、迁移和侵袭的抑制作用。结论:下调SNHG6可能通过负调控miR-186表达和调控细胞EMT过程抑制肝癌细胞的增殖、迁移和侵袭。Objective:To investigate the effect of SNHG6 on the proliferation,migration and invasion and EMT of human hepatocellular carcinoma by regulating miR-186.Methods:Real-time quantitative PCR(RT-qPCR) was used to detect the expression of SNHG6 and miR-186 in 57 liver cancer patients and hepatocellular carcinoma cells.The dual-luciferase reporter assay was used to verify the targeting regulation of SNHG6 and miR-186.Small interfering RNA of SNHG6(si-SNHG6 group),negative nonsense control sequence(si-con group),si-SNHG6 and anti-miR-186 inhibitor(si-SNHG6+anti-miR-186 group),si-SNHG6 and miR-186 inhibitor negative control(si-SNHG6+anti-miR-con group) were transfected to HepG2 cells by liposome method.MTT assay was used to detect the ability cell proliferation.The abilitities of cell migration and invasion were measured by Transwell.Western blot analysed the protein levels of CDK4,Cyclin D1, MMP-2,MMP-9 and biomakers of EMT(E-cadherin,N-cadherin,and Vimentin)in HepG2 cells.Results:Compared with normal adjacent tissues or normal liver cells,the expression of SNHG6 was increased in liver cancer patients and liver cancer cell lines,while the expression of miR-186 was decreased,showing a negative correlation.Compared with the si-con group,the activity of HepG2 cells in the si-SNHG6 group was significantly decreased,and the number of migration and invasion cells was significantly decreased,and the expression of CDK4,Cyclin D1,MMP-2,MMP-9,N-cadherin and Vimentin protein expression was decreased,and E-cadherin protein expression increased.SNHG6 targeted negative regulation of miR-186 expression,and inhibition of miR-186 expression could partially reverse the inhibitory effects of knockout of SNHG6 on proliferation,migration,invasion and EMT processes of HepG2 cells.Conclusion:Down-regulation of SNHG6 may inhibit proliferation,migration and invasion of hepatoma cells by negatively regulating miR-186 expression and regulating cell EMT processes.

关 键 词：SNHG6 肝癌 迁移 侵袭 EMT 

分 类 号：R735.7[医药卫生—肿瘤]