Evaluation of MTOR gene expression alteration during adipogenesis in human mesenchymal stem cells

Background and Aim: Obesity is a risk factor responsible for the development of several metabolic diseases. In ۲۰۱۶, the World Health Organization revealed that more than ۱.۹ billion adults, who were ۱۸ years old and older, were overweight, which over ۶۵۰ million of them were obese. Physiologically, obesity can cause by adipocyte hypertrophy and hyperplasia. Adipogenesis is the process resulting in adipose tissue hyperplasia. Different signaling pathways control the differ entiation of pre-adipocytes into adipocytes. Mammalian target of rapamycin (mTOR) is a conserved serine/threonine protein kinase that functions in two distinct complexes: mTORC۱ and mTORC۲. Recent studies show that both complexes play a crucial role in adipogenesis. It has not been known yet how MTOR gene expression changes during adipogenesis, which in this case, discovering it can help to find more effective inhibitors that are well capable of preventing and treating obesity.Methods: In this study, human mesenchymal stem cells were isolated from abdominal fat and differentiated into adipocyte under adipogenic medium. Adipogenesis was confirmed by examining the expression of PPARγ and C/EBPα genes and oil red o staining. Then the expression of PPARγ, C/EBPα and MTOR genes at ۱ day, ۵ days and ۱۰ days after differentiation was evaluated in comparison to control (undifferentiated cells) by using quantitative real-time polymerase chain reaction(qRT-PCR). Gene expression data were analyzed by one-way ANOVA in GraphPad Prism ۸ software and p-value ≤۰.۰۵ was considered significant.Results: Adipocyte differentiation has been confirmed by the results of oil red o staining and PPARγ and C/EBPα genes expression evaluation. This study showed that MTOR gene expression during adipogenesis was significantly increased by p-value ≤ ۰.۰۵. The highest MTOR gene expression was observed at five days after differentiation.Conclusion: since MTOR could be regarded as a potential therapeutic target to modulate proliferation and differentiation of adipocyte precursors, the results of the present study can help to the discovery of more effective MTOR inhibitors that can prevent and treat obesity.

Keywords:

Obesity , Adipogenesis , MTOR , Gene Expression