miR-526b-3p通过靶基因TNKS2调控肝癌细胞增殖、侵袭、迁移的分子机制探讨

Regulation of miR-526b-3p on proliferation,migration and invasion of the liver cancercell by targeting TNKS2 gene

目的:探讨miR-526b-3p对肝癌细胞增殖、迁移及侵袭的影响及其潜在的作用机制。方法:运用qRT-PCR检测人肝癌细胞HepG2、SMMC-7721、BEL-7402和正常肝细胞L02中miR-526b-3p和TNKS2的mRNA表达水平。建立miR-526b-3p过表达和TNKS2抑制表达的HepG2细胞株,采用MTT法检测细胞增殖活力,Transwell小室检测细胞的迁移及侵袭能力,Western blot检测TNKS2蛋白的表达水平;双荧光素酶报告基因分析法验证miR-526b-3p可能的靶基因。结果:与正常肝细胞L02相比,肝癌细胞HepG2、SMMC-7721及BEL-7402中miR-526b-3p的表达显著降低(P<0.05),TNKS2的表达显著升高(P<0.05)。过表达miR-526b-3p或抑制表达TNKS2均可抑制HepG2细胞的增殖、迁移及侵袭(P<0.05)。TNKS2是miR-526b-3p的靶基因。过表达TNKS2可部分逆转过表达miR-526b-3p对HepG2细胞增殖、迁移及侵袭的抑制作用。结论:miR-526b-3p可通过下调TNKS2的表达,从而抑制肝癌细胞的增殖、迁移及侵袭能力。

Objective:To explore the effect of miR-526 b-3 p on proliferation,migration and invasion of liver cancer cell and its potential mechanism.Methods:Expressions of miR-526 b-3 p in liver cancer HepG2,SMMC-7721 and BEL-7402 cells as well as normal liver cell L02 were detected by qRT-PCR.The HepG2 cell line with overexpression of miR-526 b-3 p and the HepG2 cell line with knockdown of TNKS2 were constructed.MTT,Transwell and Western blot assays were used to measure the proliferation viability,the migration and invasion cell numbers and the expression of TNKS2 protein,respectively.Possible target gene of miR-526 b-3 p was verified with dual luciferase report gene assay.Results:Compared with normal liver cell L02,the expression of miR-526 b-3 p in liver cancer cell HepG2 was significantly decreased(P<0.05),and the expression of TNKS2 was significantly increased(P<0.05).The overexpression of miR-526 b-3 p and the knockdown of TNKS2 both inhibited the proliferation,migration and invasion of HepG2 cell(P<0.05).Effect of miR-526 b-3 p on proliferation,migration and invasion of the HepG2 cell can be partially reversed by overexpression of TNKS2.Conclusion:miR-526 b-3 p can inhibit the proliferation,migration and invasion of liver cancer cells by down-regulating the expression of TNKS2.