Uba1促进C2C12细胞的分化、迁移、黏附和扩散OA

[Objective]To investigate the regulatory function of ubiquitin-like modifier activating enzyme 1(Uba1)in the growth and development of myoblasts.[Method]C2C12 myoblasts were used as a model to establish cell models with Uba1 overexpression and interference.Myogenic differentiation was assessed by immunofluorescence staining for myosin heavy chain(MyHC).Cell migration was analyzed using a wound-healing assay,and cell morphological changes were observed through cell adhesion and spreading assays.The expression of differentiation and migration marker genes was further examined by qRT-PCR and Western blotting.[Result]C2C12 cell models with stable Uba1 overexpression and interference were successfully generated.Overexpression of Uba1 significantly increased the number of MyHC-positive cells on day 5 of differentiation(P<0.01),and significantly upregulated the expression of the myogenic differentiation markers MyoG and MyHC at both the mRNA and protein levels(P<0.01).It also promoted cell migration,with a significantly increased migration distance 24 h after scratching(P<0.001)and upregulated the expression of migration-related genes Rac1,Fak,and Paxillin(P<0.05).Furthermore,Uba1 overexpression significantly increased the number of adherent cells(P<0.05)and the average cell spreading area(P<0.001).Conversely,interference of Uba1 produced the opposite effects on all the above parameters.[Conclusion]Uba1 promotes the differentiation,migration,adhesion and spreading of C2C12 cells,thereby exerting a positive regulatory role in the growth and development of skeletal muscle.