【摘要】 目的探讨微小RNA-148a (miR-148a)调控DNMT1对子宫内膜癌增殖和凋亡的影响。方法采用实时荧光定量聚合酶链反应(qRT-PCR)检测子宫内膜正常组织和癌组织中miR-148a的表达,分析miR-148a与临床指标的相关性。通过脂质体Lipofectamine 2000介导miR-148a mimics转染至子宫内膜癌细胞HEC-1B中,qRT-PCR检测转染效率,MTT和流式细胞仪检测细胞增殖凋亡情况。Transwell和Western blot检测细胞迁移侵袭能力。通过在线软件Target Scan确定DNMT1和miR-148a结合的靶位点,荧光素酶报告基因验证DNMT1和miR-148a的靶向关系。结果miR-148a在子宫内膜癌组织和细胞中低表达;过表达miR-148a抑制HEC-1B细胞增殖、迁移侵袭,促进细胞凋亡;DNMT1是miR-148a的靶基因;miR-148a抑制小鼠体内肿瘤生长和DNMT1基因表达。结论 miR-148a调控DNMT1抑制子宫内膜癌细胞增殖、迁移侵袭,促进凋亡。更多还原

【Abstract】 Objective To investigate the effect of miR-148 a targeting DNMT1 on the proliferation and apoptosis of endometrial carcinoma.Methods The expression of miroRNA-148a(miR-148 a) in normal and cancerous endometrial tissues was detected by quantitative real time polymerase chain reaction(qRT-PCR) and the correlation between miR-148a and clinical indicators was analyzed.Endometrial cancer cells HEC-1B were transfected with miR-148a mimics in vitro by Lipofectamine 2000.The transfection efficiency was detected by qRT-PCR and proliferative activity and apoptotic percentage of cells were measured by MTT and flow cytometry.Transwell and Western blot were used to detect the ability of cell migration and invasion.Target sites of DNMT1 and miR-148a were determined by online software Target Scan,and the targeting relationship between DNMT1 and mir-148a was verified by luciferase reporter gene.Results Compared with normal tissues,the expression of miR-148a in endometrial carcinoma tissues was significantly decreased.Overexpression of miR-148a inhibited the proliferation,suppressed cell migration and invasion ability,and promoted cell apoptosis of HEC-1B cells.DNMT1 is the target gene of miR-148a.miR-148a inhibited tumor growth and DNMT1 gene expression in mice.Conclusions miR-148 a regulates DNMT1 to inhibit the cell proliferation,migration and invasion ability promotes the percentage of cell apoptosis.更多还原