【摘要】 目的探讨miR-130a-3p对黑素瘤细胞的增殖、凋亡、迁移和侵袭的影响以及潜在的作用机制。方法运用实时定量聚合酶链式反应(qRT-PCR)检测黑色素瘤细胞A2058、WM239和正常人黑色素细胞HEMn中miR-130a-3p和STAT3 mRNA的表达水平;将miR-NC组(转染miR-NC)、miR-130a-3p组(转染miR-130a-3p mimics)、anti-miR-NC组(转染anti-miR-NC)、anti-miR-130a-3p组(转染anti-miR-130a-3p)、miR-130a-3p+pcDNA3.1组(共转染miR-130a-3p和pcDNA3.1)、miR-130a-3p+pcDNA3.1-STAT3组(共转染miR-130a-3p和pcDNA3.1-STAT3)、miR-NC+WT-STAT3组(共转染miR-NC和WT-STAT3)、miR-NC+MUT-STAT3组(共转染miR-NC和MUT-STAT3)、miR-130a-3p+WT-STAT3组(共转染miR-130a-3p和WT-STAT3)、miR-130a-3p+MUT-STAT3组(共转染miR-130a-3p和MUT-STAT3)均用脂质体法转染至黑素瘤A2058细胞中;采用CCK-8法检测细胞增殖;Transwell检测各组细胞迁移和侵袭;流式细胞术检测各组细胞的凋亡;Western Blot检测蛋白表达;双荧光素酶报告基因检测实验检测荧光活性。结果相较于正常人黑色素细胞HEMn,黑色素瘤细胞A2058、WM239中miR-130a-3p的表达水平显著降低(P<0.05);过表达miR-130a-3p可抑制A2058细胞增殖、迁移和侵袭,促进细胞凋亡;抑制Bcl-2、MMP-2蛋白的表达,促进Bax蛋、E-cadherin蛋白的表达。miR-130a-3p靶向调控STAT3的表达。过表达STAT3能逆转miR-130a-3p对A2058细胞增殖、迁移、侵袭的抑制和凋亡的促进作用。结论 miR-130a-3p可抑制恶性黑素瘤细胞增殖、迁移和侵袭,促进其凋亡,其机制可能与靶向下调STAT3的表达有关,将可为恶性黑素瘤的预防和治疗提供新靶点。更多还原

【Abstract】 Objective To investigate the effects of miR-130 a-3 p on proliferation,apoptosis,migration and invasion of melanoma cells,and to explore its potential action mechanism.Methods The expression levels of miR-130 a-3 p and STAT3 mRNA in melanoma cells-A2058,WM239 and normal human melanocytes-HEMn were detected by Real-time quantitative polymerase chain reaction(qRT-PCR),which in miR-NC group(transfected miR-NC),miR-130 a-3 p group(transfected miR-130 a-3 p mimics),anti-miR-NC group(transfected anti-miR-NC),anti-miR-130 a-3 p group(transfected anti-miR-130 a-3 p),miR-130 a-3 p+pcDNA3.1 group(cotransfected miR-130 a-3 p and pcDNA3.1),miR-130 a-3 p+ pcDNA3.1-STAT3 group(cotransfected miR-130 a-3 p and pcDNA3.1-STAT3),miR-NC+ WT-STAT3 group(cotransfected miR-NC and WT-STAT3),miR-NC+MUT-STAT3 group(cotransfected miR-NC and MUT-STAT3),miR-130 a-3 p+WT-STAT3 group(cotransfected miR-130 a-3 p and WT-STAT3),miR-130 a-3 p+MUT-STAT3 group(cotransformed miR-130 a-3 p and MUT-STAT3) were transfected into melanoma A2058 cells by liposome method.The cell proliferation was detected by CCK-8 method,and cell migration and invasion were detected by transwell,and cell apoptosis was detected by flow cytometry,and protein expression levels were detected by Western Blot,moreover,the fluorescence activity was detected by double luciferase reporter gene assay.Results As compared with those in normal human melanocytes-HEMn,the expression levels of miR-130 a-3 p in melanoma cells-A2058 and WM239 were significantly decreased(P<0.05).The overexpression of miR-130 a-3 p inhibited the proliferation,migration,invasion,and promoted apoptosis of A2058 cells,inhibited the expressions of Bcl-2 and MMP-2 proteins,and promoted the expressions of Bax and E-cadherin proteins.Moreover miR-130 a-3 p targeted and regulated the expression of STAT3.And the overexpression of STAT3 reversed the inhibitory effects of miR-130 a-3 p on the proliferation,migration and invasion of A2058 cells,and reversed its promotive effects on the apoptosis of A2058 cells.Conclusion The miR-130 a-3 p can inhibit the proliferation,migration,invasion of malignant melanoma cells, and promote the apoptosis of malignant melanoma cells,and its action mechanism may be related to the down-regulation of STAT3 expression,which will provide new targets for the prevention and treatment of malignant melanoma.更多还原