miR-181b通过靶定CREB1抑制人黑素瘤A375细胞的增殖和侵袭

miR-214 suppresses the growth and invasiveness of human melanoma cell line A375 by targeting CREB1

目的研究miR-181b对人黑素瘤细胞系A375的增殖与侵袭能力的影响。方法利用阳离子脂质体Lipofectamine^(TM)2000将miR-181b mimic或inhibitor转染入黑素瘤A375细胞,以使miR-181b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qRT-PCR检测核转录因子cAMP反应元件结合蛋白1(CREB1)mRNA的表达情况,采用Western blot检测CREB1蛋白的表达水平。结果与对照组相比,miR-181b沉默能够显著增强A375细胞增殖和侵袭能力,显著降低CREB1蛋白与mRNA的表达水平(P<0.01);miR-181b过表达的作用与之相反。结论 miR-181b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-181b on the proliferation and invasion of human melanoma cell line A375. Methods miR-181b mimics or inhibitor were transfeeted 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 expressions of cAMP responsive element binding protein I（CREB1） mRNA and protein were detected by qRT-PCR and Western blot respectively. Results Compared with untreated group, miR-181b silence significantly increased the proliferation and invasion ability of A375 cells and obviously decreased the expression levels of CREB 1 mRNA and protein （ P〈0.01 ） , but with opposite role for miR-181b overexpression. Conclusion The reinforcing effect of miR- 181b silence on the proliferation and invasion of A375 ceils might be related to the down-regulation of CREB1 expression.