Effect of the New Obesity-Related Gene NYGGF4 Regulate the Proliferation of 3T3-L1 Preadipocytes

Objective To investigate the effect of the new gene NYGGF4 on the proliferation of 3T3-L1 preadipocytes and its potential role in the pathogenesis of obesity.Methods The full length open read frame(ORF) of NYGGF4 was amplified by reverse transcriptase polymerase chain reaction(RT-PCR).After double digestion,the ORF was subcloned in pcDNA3.1.The NYGGF4 gene role in cell proliferation were evaluated,its eukaryotic expression vector(NYGGF4-pcDNA3.1) was constructed and transfected into 3T3-L1 preadipocytes by pcDNA3.1 transfected cells and non-transfected cells were used as a control.The stable transfected cell strain was selected with geneticin 418(G418).The mRNA level of NYGGF4 was confirmed by PCR.The stable transfected cell strain growth in 7 days was measured by microculture tetrazolium colorimetric(MTT) assay and compared to 3T3-L1 preadipocytes which was mock transfected and non-transfected 3T3-L1 preadipocytes,the growth curve was made.SPSS 10.0 software was used to analyze the data.Results 1.The stable transfected cell strain could express NYGGF4 gene;NYGGF4 mRNA was undetectable in pcDNA3.1 transfected cells.2.MTT assay showed that the proliferation rate of NYGGF4-pcDNA3.1 increased significantly than that of 3T3-L1 cell which was mock transfected and non-transfected 3T3-L1 cell.Conclusion New gene NYGGF4 can promote 3T3-L1 preadipocytes proliferation,significantly indicating that NYGGF4 gene may contribute to the development and progression of obesity.