Identification of the potential key genes for adipogenesis from human mesenchymal stem cells by RNA-Seq.

Adipogenesis, a physiological process initiated with the committed preadipocytes expressing adipocyte-specific genes and terminated in mature, differentiated and functional adipocytes, mainly involved with energy homeostasis. Abnormal distribution-changes and dysfunctions in adipogenesis may lead to complex physiopathological disorders. However, it remains unclear for the key players working for the whole complex differentiating process of adipogenesis. Here, it investigated transcriptional profiling of adipogenesis from human mesenchymal stem cells (hMSCs) by RNA-Seq transcriptome technique. Oil Red O staining assays were performed to assess adipogenic potential. Quantitative real-time PCR (qRT-PCR) and lentivirus transfection assays by small interference RNA (siRNA) were conducted to confirm the function of the candidate genes. A total of 1,078 differentially expressed genes shared at 7, 14, 21, and 28 days during adipogenesis from hMSCs, and 706 genes were significantly differentially expressed. It identified 20 potential key genes responsible for adipogenesis with four genes downregulating. The candidate gene, coagulation factor II thrombin receptor (F2R), encoding coagulation factor II thrombin receptor involving with a 7-transmembrane receptor involved in the regulation of thrombotic response, also known as proteinase-activated receptor-1, contributed to adipogenesis, especially at Day 14, by Oil Red O staining, qRT-PCR, and western blot after siRNA. A unique discovery shed new light to understand the key players of the whole processes of adipogenesis from hMSCs. The gene F2R might be used as an adipogenic marker to provide a potential target for understanding the metabolic syndromes like obesity, type-2 diabetes, steatosis, atherosclerosis, and osteoporosis.