Studys Of LncFAM200B On The Differentiation In Yak Pre-adipocytes

Yak is an excellent breed of cattle with good adaptability to the extreme environment in Qinghai-Tibet Plateau,and as the main source of production and living data to the people living in there.A tons studies indicated that fat differentiation has an important effect on improving livestock meat quality.Specifically,lncFAM200B plays an important role in fat development of Qinchuan cattle,but there are no related studies in yaks.In this study,the yak lncFAM200B was cloned and its noncoding characteristic was identified by bioinformatics and prokaryotic expression assay.The effects of lncFAM200B overexpression and interference effects on lipid metabolism-related genes,lipid droplet deposition and triglyceride content were investigated by adenovirus-mediated overexpression techniques and siRNA-mediated interference techniques in pre-adipocytes.Taken together,the effects of lncFAM200B on intramuscular adipocyte differentiation in yak were analyzed.Following are the results in this study:1.The full length of yak lncFAM200B was 531 bp,59 bp more than that in Qinchuan cattle at 83～142 bp.Bioinformatics analysis and prokaryotic expression assay showed that lncFAM200B was a real lncRNA.The results of RT-q PCR showed that the expression of lncFAM200B was higher in adipose tissue than that in muscle tissue.The lncFAM200B interaction mi RNA targets analysis revealed that many genes,such as SIRT1,SCD5,PTEN and MYPN,were related to muscle and fat development.2.The intramuscular pre-adipocytes were successfully isolated from the longissimus dorsiof yak,and then the pre-adipocytes were infected with lncFAM200B overexpressing adenovirus.The results showed that lncFAM200B overexpression significantly increased the expressions of C/EBPα,AP2(P <0.05),and lipid droplets deposition(P <0.05),and formed large lipid droplets in the cells.The triglyceride content was significantly higher than that of the control group(P <0.05)after lncFAM200B overexpression 4 days.Moreover,the SIRT1 expression trend was firstly decreased and then increased,the PTEN expression trend was firstly increased and then decreased after lncFAM200B overexpression.In addition,the cell survival rate was significantly lower than that of the control group after 3 days of lncFAM200B overexpression.In conclusion,the lncFAM200B overexpression promoted the differentiation of yak pre-adipocytes,and reduced cell proliferation efficiency.3.After the yak lncFAM200B siRNA transfected into the pre-adipocytes,the expressions of PPARγ,C/EBPα and AP2 were significantly lower than that of control group(P <0.05).Furthermore,at the later stage of differentiation,the deposition of lipid droplets in the interference group was significantly lower than that in the control group(P <0.05).The content of triglyceride was significantly lower than that of control group after lncFAM200B interference 6 days(P <0.01).During the interference of lncFAM200B,the expression trend of SIRT1 firstly increased and then decreased,whereas the expression trend of PTEN was opposite.However,the interference of lncFAM200B had no significant effects on cell proliferation.The above results showed that lncFAM200B interference inhibited the yak pre-adipocyte differentiation.In summary,lncFAM200B can promoted yak fat differentiation by increasing the expression of fat differentiation related genes C/EBPα and AP2,and the triglyceride content and the lipid droplets deposition,and lncFAM200B has important influence on the expression of target gene SIRT1,PTEN.This study laid a foundation for further exploring the mechanism of lncFAM200B in yak fat development.