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文章标题：The changes of MicroRNAs expression profiles and tyrosinase relative proteins in MITF knocked down melanocytes

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简介:The Microphthalmia associated transcription factor (MITF) is a master regulator in melanocytes proliferation, development, survival and melanoma formation. In melanocyte dysfunction disease, it¡¯s observed that the expressions of MITF, tyrosinase (TYR), tyrosinase related protein 1 (TYRP1) and tyrosinase related protein 2 (TYRP2)/dopachrome tautomerase (DCT) were changed, the consequence of which remains unclear. In this study, we focused on the change of miRNAs profiles and Tyrosinase Related Proteins (TRPs) in MITF knocked down melanocytes. For the first time, we assayed the MITF-KD microRNAs profiles using microRNA (miRNA) microarray and found that the hsa-miR-1225-3p, hsa-miR-634, hsa-miR-197, hsa-miR-766, hsa-miR-574-5p and hsa-miR-328 were up-regulated and the hsa-miR-720, hsa-miR-1308 were down-regulated in MITF knocked down melanocytes. These miRNAs were validated by miRNA real time qPCR. These miRNAs potential targets, especially the TRPs, were analyzed according to miRNA database (Sanger Center). By TargetScan predication, the hsa-miR-634 and hsa-miR-328 have poorly conserved site on TYR and hsa-miR-197 have poorly conserved site on TYR1. Through qPCR and western blot we found that the expression of TYR and TYRP1 were dramatically decreased and the expression of TYRP2 was increased in MITF knocked down melanocytes (MITF-KD). These results implicated that the miRNAs may involved in MITF regulation on TYR, TYRP1 and TYRP2 which provide a new clue for understanding the role of miRNAs in melanocyte dysfunctional disease.