A Study on the Mechanism of miR-126 on the Migration and Invasion of Cutaneous Melanoma C8161
This study focused on the effects of the differential expression of microRNA-126-3p(miR-126)on migration and invasion of human cutaneous melanoma(CM)cell line C8161,and the role of Janus tyrosine protein kinase 2/signaling and transcriptional activator 3(JAK2/STAT3)pathway and epithelial-mesenchymal transition(EMT)process were investigated.Dysregulation of miR-126 was achieved by cell transfection,and miR-126-deregulated cells were treated with the JAK2/STAT3 pathway inhibitor AG490 or the agonist Coumermycin A1.Thus,C8161 cells were divided into 5 groups:control group(without transfection or drug treatment),negative control(NC)group(with mimics control transfection,but no drug treatment),miR-126 group(with miR-126 mimics transfection,but no drug treatment),miR-126+pathway inhibitor group(with miR-126 mimics transfection and AG490 treatment),miR-126+pathway agonist group(with miR-126 mimics transfection and Coumermycin A1 treatment).Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression level of miR-126.The expression levels of EMT key proteins and JAK2/STAT3 pathway proteins were determined by Western blotting.Cell viability,migration and invasion abilities were determined by cell counting kit-8(CCK-8),scratch healing assay and Transwell cham-ber combined with matrix gel,respectively.Compared with the NC group,transfection with miR-126 mimics significantly in-creased the expression level of miR-126 in the miR-126 group(#P<0.05),while the relative cell vitality,cell migration rate and cell invasion number significantly decreased(#P<0.05),accompanied with higher protein level of E-cadherin(#P<0.05),and lower protein levels of Vimentin,N-cadherin,fibronectin(FN),JAK2,STAT3,p-JAK2,and p-STAT3,and as well as lower ratios of p-JAK2/JAK2 and p-STAT3/STAT3(#P<0.05).More importantly,compared with NC group and miR-126 group,the above indicators of JAK2 and STAT3 protein levels were further changed in miR-126+pathway inhibitor group(#P<0.05 and&P<0.05),whereas all the above indexes were less changed in miR-126+pathway activator group(#P<0.05 and&P<0.05),and cell invasion number and FN,JAK2 and STAT3 protein levels in miR-126+pathway activator group were little different from NC group without significance.Overexpression of miR-126 inhibits cell viability,migration and invasion of human CM cells presumably by blocking EMT process and inhibiting the activation of JAK2/STAT3 pathway.These results were helpful to provide a new theoretical basis for the clinical treatment of human CM and a new strategy for its treatment.
cutaneous melanomamicroRNA-126-3pJanus tyrosine protein kinase 2/signal transduction and transcription activator 3migrationinvasion
国家科技期刊平台
NSTL
万方数据
维普
