Relationship between expression levels of RPS۶KB۱ and GLI۱ with DYRK۱B during adipogenesis of ADSCs in the presence and absence of R۱۰۲C mutation in DYRK۱B gene

Background and Aim: Metabolic syndrome is increasing as a worldwide health problem, including Iran. The missense mutation of the DYRK۱B gene R۱۰۲C in an Iranian population has been shown to be correlated to a rare autosomal-dominant form of the metabolic syndrome. Rate of adipogenesis is increased in the presence of the mutated gene which can be through the interactions with Shh pathway although the details of DYRK۱B interactions with other components of adipogenic pathways is not completely understood. Gli۱ is a transcription factor and is the final effector of Shh pathway.Gene RPS۶KB۱ encodes a kinase called S۶K and plays a role at early stages of adipogenesis.C/EBPα and PPARγ are two major regulators of adipogenesis and their expression levels increased during this process. In the current study we assessed the expression levels of DYRK۱B, GLI۱,RPS۶KB۱ , C/EBPα and PPARγ in the day۱, ۵ and ۱۰ after adipogenesis induction .Methods: The ADSCs were obtained from two groups of donors, affected with R۱۰۲C mutation in DYRK۱B gene and non-affected ones, characterized using flow cytometry. The obtained ADSCs were cultured and differentiated into adipocytes. Adipogenic differentiation was confirmed with oil Red o staining. After total RNA extraction and cDNA synthesis, the gene expression levels were evaluated by Real-time PCR. The results were analyzed by two-way ANOVA test using Graph Pad Prism.Results: After adipogenesis induction the expression of C/EBPα and PPARγ in both groups increased, also they were higher in the mutated group significantly. DYRK۱B expression increased during the day۱ and ۵ and decreased at day۱۰, not analytically significant. RPS۶KB۱ increased in the day۱ and ۵ and decreased at day۱۰; GLI۱ expression decreased in all days. Although Changes of RPS۶KB۱ and GLI۱ were significant within each group, it wasn’t between the groups.Conclusion: The R۱۰۲C mutation of DYRK۱B can increase adipogenesis. During adipogenesis, Shh pathway activity is decreased (according to GLI۱ descending levels). RPS۶KB۱ ascending levels in day۱ and ۵ is correlated with early stages of adipogenesis. Since the changes of RPS۶KB۱ and GLI۱ aren’t significant between groups, different rates of adipogenesis in the case R۱۰۲C mutation cannot be related to GLI۱ or S۶K directly.

Keywords:

Metabolic Syndrome , adipogenesis , DYK۱B , GLI۱ , RPS۶KB۱ , S۶K