Influence of silencing the MC4R gene by lentivirus-mediated RNA interference in bovine fibroblast cells

摘要

Melanocortin receptor 4 (MC4R) is a key element in the mechanisms used to regulate both aspects of keeping the balance between energy uptake and energy expenditure.?MC4R?was knocked down by lentivirus-mediated shRNA expressing plasmids, which were controlled by the U6 promoter in bovine fibroblast cells, and the expression of?MC4R?was examined by the real time-PCR and Western blot analysis.? We further used Real time-PCR analysis was used to characterize the expression of Leptin,?POMC,?AGRP,?MC3R?and?NPY?gene. The relative genes [leptin, proopiomelanocortin (POMC), agouti-related peptide (AGRP),?MC3R?and neuropeptide Y (NPY)] expression level seemed to be closely associated with theMC4R?gene in bovine fibroblast cell lines (BFCs). The levels of both?MC4R?mRNA and protein were significantly reduced by RNA interference (RNAi) mediated knockdown of?MC4R?in BFCs cells transfected with plasmid-based?MC4R-specific shRNAs. The finding of this study demonstrated that vector based siRNA expression systems were an efficient approach to the knockdown of the?MC4Rgene expression in bovine fibroblast cells and they provided a new molecular basis for understanding the relationship of?MC4R?and other genes, which were responsible for the regulation of energy homeostasis by the melanocortin system.