miR-182靶向作用環(huán)磷酸腺苷反應元件結合蛋白1調控人黑素瘤A375細胞的增殖和侵襲
發(fā)布時間:2018-03-25 20:37
本文選題:miR- 切入點:增殖 出處:《解剖學報》2017年06期
【摘要】:目的探討miR-182對人皮膚黑素瘤細胞系A375細胞的增殖與侵襲能力的影響,及miR-182影響黑色素細胞增殖的機制。方法利用陽離子脂質體LipofectamineTM2000將miR-182模擬物或抑制劑轉染人黑素瘤A375細胞,以使miR-182過表達或低表達,采用MTT方法檢測細胞活力變化,應用Transwell小室法檢測A375細胞侵襲能力的變化,同時通過Real-time PCR檢測核轉錄因子環(huán)磷酸腺苷反應元件結合蛋白1(CREB1)mRNA的表達情況,采用Western blotting檢測CREB1蛋白的表達水平。結果與對照組相比,miR-182沉默能夠顯著增強A375細胞增殖和侵襲能力,P0.01,顯著降低CREB1蛋白與mRNA的表達水平,P0.01;miR-182過表達的作用與之相反。結論 miR-182沉默增強A375細胞增殖和侵襲能力可能與降低CREB1蛋白相關。
[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.
【作者單位】: 福建醫(yī)科大學附屬泉州第一醫(yī)院皮膚科;沈陽市第七人民醫(yī)院皮膚科;
【分類號】:R739.5
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