Transcriptome Analysis of C2C12 Cells Induced to Differentiate into Adipocytes
TIAN Hongwei1,2, Meirong1,2, LI Yuling1,2, Wuyundalai1,2*
To explore the key genes in the transdifferentiation of C2C12 cells into adipocytes, the medium supplemented with IBMX, dexamethasone, insulin and rosiglitazone was used to induce C2C12 cells for six days. The differentiation of cells before and after induction was identified by oil red O staining and immunofluorescence staining. Transcriptome analysis and RT-qPCR (real-time fluorescence quantitative PCR) techniques were used to evaluate the expression levels of differentially expressed genes; Western blot was employed to measure the protein levels; and ELISA (enzyme-linked immunosorbent assay) was applied to analyze the cell state. The results showed that after six days of induction, a large number of lipid droplets appeared after oil red O staining of C2C12 cells, and a positive signal of FABP4 was detected by immunofluorescence staining. GO (Gene Ontology) analysis and KEGG (Kyoto Encyclopedia of Genes and Genomes) pathway analysis revealed that the expression levels of genes related to viral response and immune system pathways were significantly upregulated. The Hub genes Isg15, Ddx58, Ifit3, Irgm2, Eif2ak2, Irf9 and Stat1 were further screened. RT-qPCR verified the up-regulated expression of Isg15, Fabp4 and C/EBPα expression, and immunoblot confirmed the increase of FABP4 and IL-6 protein levels. Upregulated expression of Ddx58, Ifit3, Irgm2, Eif2ak2, Irf9, and Stat1 genes were involved in the inflammatory response to maintain cell survival. Taken together, this study revealed part of the molecular mechanism of myocyteto-adipocyte transformation, providing a basis for further understanding of myogenic adipogenesis.
CN
EN