miR-146a对人黑色素瘤A375细胞增殖和侵袭的影响及作用机制

Effect and mechanism of miR-146a on proliferation and invasion of human melanoma A375 cells

目的探讨微小RNA-146a(miR-146a)对人黑色素瘤A375细胞增殖与侵袭能力的影响及可能机制。方法利用阳离子脂质体LipofectamineTM2000将miR-146a模拟物或抑制剂转染入黑色素瘤A375细胞,使miR-146a过表达或低表达,采用MTT检测细胞活力变化,应用Transwell小室法检测A375细胞侵袭能力的变化。根据生物信息学软件预测FBXL10为miR-146a的下游靶基因,双荧光素酶报告分析进行验证。分别采用实时qPCR与Western blotting检测转染后各组A375细胞FBXL10 mRNA与蛋白的表达水平。结果与对照组比较,miR-146a模拟物能够显著抑制A375细胞增殖和侵袭能力(均P<0.05)。双荧光素酶报告分析结果显示,FBXL10为miR-146a的下游靶基因,miR-146a过表达可显著降低FBXL10蛋白与mRNA的表达水平;miR-146a抑制剂的作用则与之相反。结论miR-146a过表达能够抑制A375细胞增殖和侵袭能力,其作用机制可能与降低FBXL10表达相关。

Objective To investigate the effect and mechanism of microRNA-146a(miR-146a)on proliferation and invasion of A375 cells.Methods miR-146a mimics or inhibitors were transfected into A375 cells using a lipofectamine package.MTT assay was used to evaluate A375 cell viability and Transwell chamber assay was used to measure invasion.According to bioinformatics analysis,FBXL10 was predicted to be the downstream target gene of miR-146a,which was validated by double luciferase reporter analysis.FBXL10 mRNA and protein expression levels were detected by RT-qPCR and Western blotting,respectively,after transfection with miR-146a mimics or inhibitors.Results Compared to the miR-146a negative control group,miR-146a mimics could significantly inhibit the A375 cells invasion and proliferation(P<0.05).Double luciferase reporter analysis showed that FBXL10 was the downstream target gene of miR-146a.miR-146a mimics significantly reduced FBXL10 protein and mRNA expression,whereas miR-146a inhibitors had the opposite effect.Conclusion miR-146a mimics could inhibit proliferation and invasion of A375 cells and the mechanism might be related to reduced FBXL10 expression.