Glycoprotein M6B knockdown promotes white adipocytes to brown adipocyte phenotype

  AIM  To investigate the role of glycoprotein M6B (GPM6B) in phenotypic transformation of white adipocytes to brown adipocytes.

  METHODS  The expressions of GPM6B in white adipose tissue and brown adipose tissue were detected by Western Blot and qPCR. The C3H10T1/2 cells were infected with lentiviruses expressing two independent hairpin shRNA sequences for GPM6B (shGPM6B1 and shGPM6B2) or control shScramble sequence. The transfection efficiency of lentiviruses was observed by fluorescence microscope. The extent of the knockdown by the targeted shRNAs for GPM6B was confirmed by Western blot and qPCR analysis. The control (shScramble) and GPM6B knockdown (shGPM6B1 or shGPM6B2) stable cell lines were treated to induce adipogenesis for 8 days and examined for lipid accumulation by oil red O staining. The protein levels of UCP1 were tested by Western blot and the mRNA levels of the thermogenetic genes were tested by qPCR.

  RESULTS  Compared with that in WAT, the expression of GPM6B was significantly down-regulated in BAT. Each of the shRNA constructs effectively knocked down the mRNA and protein expression of GPM6B (P<0.05). Remarkably smaller lipid droplets were observed in differentiated C3H10T1/2 cells when GPM6B was inhibited. Moreover, GPM6B knockdown significantly promoted the expression of UCP1 and the thermogenetic genes during this process (P<0.05).

  CONCLUSION  GPM6B knockdown promotes phenotypic transformation of white adipocytes to brown adipocytes.