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,P<0.01,显著降低CREB1蛋白与mRNA的表达水平,P<0.01;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。
Objective To investigate the effects of miR-182 on the proliferation and invasion of human melanoma cell line A375 and to elucidate its action mechanisms. Methods miR-182 mimics or inhibitor were transfected into A375 cells by lipofectamine package, MTT was applied to detect the viability of A375 cells, transwell chamber assay was used to measure the invasion of A375 cells, the expression levels of cyclic adenosine monophosphate responsive element binding protein 1 ( CREB1 ) mRNA and protein were detected by Real-time PCR and Western blotting respectively. Results Compared with the untreated group, miR-182 silence significantly increased the proliferation and invasion ability of A375 cells and obviously decreased the expression levels of CREB1 mRNA and protein, P 〈 0. 01, but with opposite role for miR- 182 overexpression. Conclusion The reinforcing effect of miR-182 silence on the proliferation and invasion of A375 ceils may be related to the down-regulation of CREB1.
出处
《解剖学报》
CAS
CSCD
北大核心
2017年第6期710-714,共5页
Acta Anatomica Sinica