Mature miR-183, negatively regulated by transcription factor GATA3, promotes 3T3-L1 adipogenesis through inhibition of the canonical Wnt/β-catenin signaling pathway by targeting LRP6

Differentiation of preadipocytes into adipocytes and the formation of the subsequent adipose tissue are critical for mammalian growth and development. The molecular mechanism relating to preadipocyte differentiation and adipogenesis from the perspective of miRNAs is not yet completely understood. Here we investigated whether miR-183 functioned in the differentiation process. Both gain-of-function and loss-of-function assays demonstrated that miR-183 positively regulated 3T3-L1 differentiation by enhancing the expression of adipogenic marker genes such as CCAAT/enhancer binding protein α (C/EBPα), peroxisome proliferator-activated receptor γ (PPARγ), adiponectin and fatty acid synthase (FAS), as well as the triglyceride content and accumulation of lipid droplets. Meanwhile, low-density lipoprotein receptor-related protein 6 (LRP6) was known to impair the canonical Wnt/β-catenin signaling pathway and thereafter reduce c-myc and nuclear β-catenin protein. We showed that the inhibition of LRP6 by siRNA promoted 3T3-L1 adipogenic differentiation and adipogenesis. Further analysis showed that mouse miR-183 gene had its own transcription unit containing CpG islands, transcription start site (TSS), coding sequence (CDS) and polyA signal within the flanking sequences 2500 nt upstream and downstream of mouse miR-183 in genome. The core promoter of miR-183 gene was identified and transcription factor GATA3 (GATA binding protein 3) significantly inhibited the expression of mature miR-183 by binding to its core promoter in vivo, as indicated by thechromatin immunoprecipitation (ChIP) assay. These results suggest that miR-183, though negatively regulated by transcription factor GATA3, enhances 3T3-L1 preadipocyte differentiation and adipogenesis through the inhibition of the canonical Wnt/β-catenin signaling pathway by targeting LRP6.