miR-182靶向作用环磷酸腺苷反应元件结合蛋白1调控人黑素瘤A375细胞的增殖和侵袭

发布时间：2018-03-25 20:37

本文选题：miR-　切入点：增殖　出处：《解剖学报》2017年06期

【摘要】：目的探讨miR-182对人皮肤黑素瘤细胞系A375细胞的增殖与侵袭能力的影响,及miR-182影响黑色素细胞增殖的机制。方法利用阳离子脂质体LipofectamineTM2000将miR-182模拟物或抑制剂转染人黑素瘤A375细胞,以使miR-182过表达或低表达,采用MTT方法检测细胞活力变化,应用Transwell小室法检测A375细胞侵袭能力的变化,同时通过Real-time PCR检测核转录因子环磷酸腺苷反应元件结合蛋白1(CREB1)mRNA的表达情况,采用Western blotting检测CREB1蛋白的表达水平。结果与对照组相比,miR-182沉默能够显著增强A375细胞增殖和侵袭能力,P0.01,显著降低CREB1蛋白与mRNA的表达水平,P0.01;miR-182过表达的作用与之相反。结论 miR-182沉默增强A375细胞增殖和侵袭能力可能与降低CREB1蛋白相关。
[Abstract]:Objective to investigate the effect of miR-182 on the proliferation and invasion of human skin melanoma cell line A375 and the mechanism of miR-182 affecting the proliferation of melanoma cell. Methods miR-182 mimics or inhibitors were transfected into human melanoma A375 cells by cationic liposome LipofectamineTM2000. In order to make miR-182 overexpression or low expression, MTT method was used to detect cell viability, Transwell chamber assay was used to detect the invasion ability of A375 cells, and Real-time PCR was used to detect the expression of nuclear transcription factor cyclic adenosine monophosphate binding element binding protein 1(CREB1)mRNA. Western blotting was used to detect the expression of CREB1 protein. Results compared with the control group, the silencing of pmiR-182 could significantly enhance the proliferation and invasion ability of A375 cells and significantly reduce the expression level of CREB1 protein and mRNA. It is suggested that miR-182 silencing enhances the proliferation and invasion of A375 cells, which may be related to the reduction of CREB1 protein.
【作者单位】：福建医科大学附属泉州第一医院皮肤科;沈阳市第七人民医院皮肤科;
【分类号】：R739.5

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