Expression and molecular regulation of key genes during adipogenesis of neural crest stem cells

Abstract

ABSTRACTNeural crest stem cells (NCSCs) are pluripotent stem cells derived from the “fourth germ layer”. Similar to mesenchymal stem cells (MSCs), NCSCs can differentiate into a variety of cell types, such as adipocytes. However, the mechanism of NCSCs adipogenesis remains unclear. Previously, we have revealed that primary cells have neural crest and stem cell properties and can differentiate into adipocytes. Therefore, in this study, the differentially expressed genes (DEGs) of NCSCs at specific time points of adipogenesis were predicted by mRNA sequencing, the key genes of adipogenesis were predicted by principal component analysis, heat map, GO and KEGG enrichment analysis, and the expression of DEGs was verified by RT-qPCR. RNA interference (RNAi) technology was used to inhibit the expression of DEGs, and RT-qPCR and Western blot were used to explore the regulatory mechanism between DEGs. Sequencing results indicated a possible regulatory relationship between C/EBPα, C/EBPβ, C/EBPδ and PPARγ. The results of RT-qPCR were consistent with those of mRNA sequencing. Combined with RT-qPCR and western blot results, we found that C/EBPβ and PPARγ regulated the transcription of C/EBPα during NCSCs adipogenesis, and C/EBPβ and PPARγ formed positive feedback loop.