Screening and validation of downstream target genes of SLC25A21 based on bioinformatics

Solute carrier 25 member 21 (SLC25A21) serves as an oxodicarboxylate carrier, which mainly conveys 2-oxoadipate from the cytoplasm to the mitochondria via a reverse exchange mechanism. Previous studies have indicated that the capacity for glucose consumption is significantly enhanced in 3T3-L1 cells overexpressing SLC25A21. In this study, we upregulate SLC25A21 in 3T3-L1 cells to further probe into the downstream key metabolic genes of SLC25A21. Through high-throughput sequencing combined with bioinformatics analysis, differentially expressed genes are obtained, and the expression of key genes is verified by qRT-PCR. The results demonstrat that: (1) There are 26 up-regulated genes and 66 down-regulated genes in the adipocytes overexpressing SLC25A21; (2) GO (gene ontology) analysis indicates that the biological functions of differentially expressed genes are predominantly involved in lipid synthesis and metabolism, and KEGG (Kyoto encyclopedia of genes and genomes) and GSEA (gene set enrichment analysis) analyses reveal that differentially expressed genes are mainly concentrated in sphingolipid metabolism, secretion and synthesis of insulin and glucagon-like peptide 1; (3) By means of cytoHubba, 10 key genes with the highest scores, such as GRB2, SOS1, SHC1, CBL, HRAS, SOS2, EGFR, MET, PLCG2 and KRAS, were screened out and they are mainly involved in the sugar and lipid metabolism processes of cells; (4) SLC25A21 is overexpressed in adipocytes, and the qRT-PCR verification results show that the mRNA expression levels of other genes increased correspondingly, except for KRAS expression, which exhibits no significant change. These results provide a theoretical basis for further investigations on the role and mechanism of SLC25A21 in the process of glucose and lipid metabolism.