Suppression of MafA mRNA with siRNA prevents adipose cell differentiation in 3T3-L1 cells.

摘要

One of the large Mafs, MafA protein, is a strong transactivator of insulin in pancreatic beta cells. Mafs are also known to play important roles in a variety of developmental and differentiation processes in many organs and tissues. Adipocytes are highly involved in insulin actions and glucose and lipid metabolism, and their proliferation and differentiation is regulated by coordination of several signal transduction and transcriptional factors, including members of the Maf family. To explore the role of MafA in adipocytes, we modified the MafA mRNA level in cultured adipocytes by the RNA interference technique and analyzed the resulting morphological changes and changes in expression of related genes. MafA siRNA was transfected into 3T3-L1 adipocytes. Expression of MafA was confirmed by real-time PCR and Western blotting. Expression of adipocytokines and transcriptional factors was also measured by real-time PCR. Cells were examined for morphological changes and lipid accumulation by microscopy. The MafA expression level in the MafA-siRNA-transfected pre-adipocytes was reduced by approximately 30% on day 0 pre-induction and by approximately 70% on day 3 post-induction, in comparison with stop-siRNA-transfected cells. Cell growth and lipid droplet accumulation were prevented by MafA mRNA suppression, and peroxisome proliferator-activated receptor (PPAR) gamma2 and CCAAT/enhancer-binding proteins (C/EBP)alpha, both of which are transcriptional factors essential for adipocyte differentiation, were down-regulated. Expression of the genes encoding the adipocytokines, adiponectin and adipsin was also suppressed. The results suggested a possible role of the transcriptional factor MafA in regulation of adipocyte function and differentiation.