MiR-503靶向bcl-2增强BEL-7402细胞对顺铂的敏感性

摘要

目的:探讨miR-503是否能通过调控bcl-2的表达增强BEL-7402细胞对顺铂的敏感性.方法:采用实时荧光定量PCR检测肝癌细胞中miR-503和bcl-2 mRNA的表达水平;Western印迹观察肝癌细胞中Bcl-2蛋白的表达水平;脂质体转染法将miR-503模拟物瞬时转染至BEL-7402细胞;生物信息学软件预测miR-503潜在靶基因;双荧光素酶活性验证miR-503潜在的靶位点;MTT法检测细胞药物敏感性的改变.结果:相比于HL-7702细胞,miR-503在BEL-7402细胞中表达水平下调,Bcl-2蛋白的表达水平上调;miR-503能够与bcl-2靶向结合,下调bcl-2的表达;miR-503转染组的细胞活力较miRNA阴性对照组显著降低.结论:MiR-503可能通过靶向bcl-2增强BEL-7402细胞对顺铂的药物敏感性,抑制肝癌细胞增殖.%Objective:To investigate effects of miR-503 on cisplatin sensitivity in BEL-7402 cells by targeting of bcl-2.Methods:MiR-503 and bcl-2 mRNA expression levels in hepatocellular carcinoma cells were measured by real-time quantitative (qRT)-PCR;Bcl-protein level was detected by Western blot;miR-503 mimics were transiently transfected to the BEL-7402 cells by liposome transfection;potential target genes of miR-503 were predicted by Bioinformatics software;miR-503 potential targets were validated by dual luciferase activity;and the cell viability was measured by MTT assay.Results:MiR-503 level was down-regulated and Bcl-2 protein expression level was up-regulated in BEL-7402 cells compared with HL-7702 cells.MiR-503 could interact with bcl-2 and inhibit its expression.Cell vitality with miR-503 transfection was significantly reduced compared to that in the negative control.Conclusion:MiR-503 may enhance the sensitivity of BEL-7402 cells to cisplatin and inhibit the cell proliferation by targeting bcl-2.