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作 者:杨晓燕[1,2] 殷杰[1] 向琼[3] 谢红艳[1] 虞佳[3] 甘润良[2,4] 雷小勇[2,3]
机构地区:[1]南华大学生物研究所,湖南衡阳421001 [2]南华大学湖南省分子靶标新药研究协同创新中心,湖南衡阳421001 [3]南华大学药物药理研究所,湖南衡阳421001 [4]南华大学肿瘤研究所,湖南衡阳421001
出 处:《中南大学学报(医学版)》2017年第6期605-610,共6页Journal of Central South University :Medical Science
基 金:国家自然科学基金(81372579);湖南省教育厅科学研究项目(17C1402;14C0998);湖南省研究生创新项目(CX2015B386)~~
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(q RT)-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.
关 键 词:MiR-503 BCL-2 BEL-7402细胞 顺铂
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