miR-485-5p靶向作用于SOX5抑制肝癌细胞Hep3B的活力及侵袭和迁移能力

miR-485-5p inhibits viability and migration and invasion abilities of hepatocellular carcinoma Hep3B cells by targeting SOX5

目的:研究微小RNA-485-5p(miR-485-5p)对肝癌细胞的活力及迁移和侵袭能力的影响及其潜在机制。方法:以正常肝细胞THLE-3为对照,采用RT-qPCR检测肝癌细胞中性别决定区Y框蛋白5(SOX5)mRNA和miR-485-5p的表达,Western blot检测SOX5、增殖细胞核抗原(PCNA)、Ki67、细胞周期蛋白D1(cyclin D1)及基质金属蛋白酶2(MMP-2)的表达水平,MTT法检测Hep3B细胞的活力,Transwell实验检测细胞的迁移和侵袭能力,双萤光素酶报告基因系统验证细胞中miR-485-5p与SOX5的调控关系。结果:与对照组相比,在肝癌细胞Hep3B、Huh7和HCCLM3中miR-485-5p的表达水平显著降低(P<0.05),SOX5的mRNA和蛋白表达水平均显著升高(P<0.05);过表达miR-485-5p可抑制Hep3B细胞的活力及迁移和侵袭能力(P<0.05);miR-485-5p靶向调控SOX5的表达,敲减SOX5的表达也可抑制Hep3B细胞的活力及迁移和侵袭能力;过表达SOX5可部分逆转过表达miR-485-5p对Hep3B细胞活力及迁移和侵袭能力的抑制作用(P<0.05)。结论:miR-485-5p通过靶向调控SOX5基因抑制Hep3B细胞的活力及迁移和侵袭能力,有望成为肝癌诊断和治疗的潜在分子靶点。

AIM:To investigate the effects of microRNA-485-5p(miR-485-5p)on the viability,migration and invasion abilities of hepatocellular carcinoma cells and the underlying mechanism.METHODS:The expression levels of sex determining region Y-box 5(SOX5)mRNA and miR-485-5p in the hepatocellular carcinoma Hep3B cells were detected by RT-qPCR with normal hepatocyte THLE-3 as control.Western blot was used to measure the expression levels of SOX5,proliferating cell nuclear antigen(PCNA),Ki67,cyclin D1 and matrix metalloproteinase-2(MMP-2).The viability of Hep3B cells was measured by MTT assay.The migration and invasion abilities of the Hep3B cells were detected by Transwell assay.Dual-luciferase reporter assay system was applied to verify the relationship between miR-485-5p and SOX5.RESULTS:Compared with the control cells,the expression level of miR-485-5p was decreased in hepatocellular carcinoma cells Hep3B,Huh7 and HCCLM3(P<0.05),while the expression of SOX5 at mRNA and protein levels were significantly increased(P<0.05).Over-expression of miR-485-5p inhibited the viability,migration and invasion of Hep3B cells.miR-485-5p targeted the 3′-UTR of SOX5 and negatively regulated the expression of SOX5.Knocking-down of SOX5 expression inhibited the viability,migration and invasion of Hep3B cells.Over-expression of SOX5 partially reversed the inhibitory effect of miR-485-5p over-expression on the viability,migration and invasion of Hep3B cells.CONCLUSION:miR-485-5p inhibits the viability,migration and invasion of Hep3B cells by targeting SOX5 gene.miR-485-5p is a potential molecular target for hepatocellular carcinoma.