The Role Of SHOX2 In Adipogenesis Differentiation Of Human Adipose Stem Cells

Adipose tissue is one of the largest tissue in the human body,there are mainly two kinds of forms:white adipose tissue(WAT)and brown adipose tissue(BAT).Obesity is a very serious health problem which has a relationship with the formation of fat.The increase in the number of fat cells and the enlargement of the fat cell volume are the main reasons for the formation of obesity.The mature fat cells mainly come from the differentiation of adipose stem cells,so there is a great significance to know the differentiation process of the adipose stem cells.The process of fat formation is regulated by a series of related factors.Peroxisome proliferator-activated receptor ?(PPARy)and CCAAT/enhancer-binding protein ?(C/EBP?)are the main functions.BMP and Wnt signaling pathways are also involved.SHOX2 is a member of homeo-domain transcription factors that has an important function in the development of bones and heart,and it also has a relationship with BMP signaling pathway.The studies have shown that in rodents the expression of Shox2 is higher in the subcutaneous fat than the visceral fat.Shox2 can control the rate of lipolysis through the regulation of the expression of Adrb3,but now the information about SHOX2 on the effects of adipogenesis differentiation is very rare,so this study focuses on whether SHOX2 has a role in the differentiation in adipogenesis differentiation,and then provide further data network of adipogenesis differentiation and the occurrence of obesity.Firstly,several kinds of stem cells include human adipose stem cells,human bone marrow mesenchymal stem cells and human dental pulp stem cells were isolated,and then induced in vitro into mature adipose cells.Detecting the expression of various isoforms of SHOX2 before and after the differentiation.The results demonstrate that these isoforms is always there in the process of the adipogenesis before and after the differentiation especially the SHOX2b.Then we used the SHOX2b as the targeted gene to construct LentiCRISPRv2-SHOX2 knockout plasmid by CRISPR/Cas9-mediated genome engineering,at the same time we construct pCEP4-SHOX2 overexpression plasmid.The plasmids were respectively used with liposome transfected into the human adipose stem cells.The results showed that the most of the adipose stem cells have fomation into fat by overexpression of SHOX2,but rale fat were taken shape by knock-out SHOX2,so SHOX2 play a promoting role in adipogenesis differentiation.In summary,SHOX2 is an important factor in adipogenesis differentiation of human adipose stem cells.Our experimental results provided the relevant experimental data in the fat development and the occurrence of obesity.