miR-548c-3p通过调控TRIM59表达对肝癌细胞增殖、侵袭和迁移的影响

miR-548c-3p inhibits cell proliferation, migration and invasion of hepatocellular carcinoma HepG2 cells by targeting TRIM59

目的研究miR-548c-3p对肝癌细胞增殖、迁移和侵袭的影响和潜在的分子机制。方法 qRT-PCR检测TRIM59 mRNA和miR-548c-3p的表达,Western blot检测TRIM59、增殖相关蛋白CyclinD1和p21,及迁移侵袭相关蛋白MMP2和MMP9的表达水平,MTT法测定HepG2细胞增殖活性,Transwell实验检测细胞迁移和侵袭能力,双荧光素酶报告系统验证miR-548c-3p与TRIM59的调控关系。结果与正常肝细胞L02相比,在肝癌细胞HepG2、SMMC-7721和BEL-7402组中TRIM59的mRNA和蛋白表达量均显著升高(P<0.05),miR-548c-3p的表达量则均显著降低(P<0.05);过表达miR-548c-3p和抑制TRIM59表达均可抑制HepG2细胞增殖、迁移和侵袭;双荧光素酶报告系统结果表明,miR-548c-3p靶向负调控TRIM59的表达;过表达TRIM59逆转了过表达miR-548c-3p对HepG2细胞增殖、迁移和侵袭的抑制作用。结论 miR-548c-3p通过靶向TRIM59基因抑制HepG2细胞的增殖、迁移和侵袭,miR-548c-3p是肝癌的潜在治疗靶点。

Objective To investigate the efffect of miR-548c-3p on proliferation,migration and invasion of hepatocellular carcinoma cells and the underlying mechanism. Methods qRT-PCR was used to measure the expression levels of TRIM59 mRNA and miR-548c-3p. Western blot was applied to detect the expression levels of TRIM59,proliferation-related proteins CyclinD1 and p21,as well as the migration and invasion related proteins MMP2 and MMP9. MTT assay was carried out to detect the proliferation of HepG2 cells. Transwell assay was carried out to determine the migration and invasion ability of HepG2 cells. Dualluciferase reporter assay was implemented to verify the relationship between miR-548c-3p and TRIM59.Results Compared with human normal hepatocytes L02,the expression levels of TRIM59 mRNA and protein were significantly increased in hepatocellular carcinoma cells HepG2,SMMC-7721 and BEL-7402(P<0.05),while the expression level of miR-548c-3p was remarkably decreased(P<0.05). Overexpression of miR-548c-3p and inhibition of TRIM59 inhibits the proliferation,migration and invasion of HepG2 cells. The results of dual-luciferase reporter assay showed that miR-548c-3p targeted and negatively regulated the expression of TRIM59. Overexpression of TRIM59 reversed the inhibitory effect of overexpression of miR-548c-3p on the proliferation,migration and invasion of HepG2 cells. Conclusion miR-548c-3p inhibits the proliferation,migration and invasion of HepG2 cells by targeting TRIM59 gene,a potential therapeutic target for hepatocellular carcinoma.