GIT1 promotes osteoblast migration by regulating ERK1/2 activity in focal adhesions / 中华创伤骨科杂志

ABSTRACT

Objective To study the function and mechanism of GIT1(G protein coupled receptor kinase interacting protein 1)in osteoblast migration.Methods GIT1 and ERK1/2(Extracellular Signal-regulated ki- nase 1/2)were detected in mice primary osteoblasts.The localizations of GIT1 and ERK1/2 were determined by immunofluorescence stain with or without PDGF(platelet-derived grnwth factor)stimulation.The association of GIT1 anti ERK1/2 was analyzed by co-immunoprecipitation and western blot.After stimulation,the co-localization of GIT1 and pERK1/2 in osteoblasts was detected by double-immunnfluorescence stain.The pERK1/2 localization was detected by immunofluorescence stain after GIT1RNAh adenovirus infection of osteoblasts.The role of this associa- tion was determined by wound healing assay.Results The co-immunoprecipitation results showed that GIT1 in- teracted with ERK1/2 in osteoblasts induced by PDGF and this association occurred in focal adhesions.GIT1 RNAh adenovirus significantly inhibited the pERK1/2 translocation to focal adhesions and osteoblast migration induced by PDGF.Conclusion GIT1 associates with ERK1/2 in osteoblasts,which is required for pERK1/2 translocation to focal adhesions and osteoblast migration.