Expression of Halo-hFGF18 and study of its effect on differentiation of ATDC5 cells

Fibroblast growth factor 18 (FGF18) is a member of the fibroblast growth factor family and important in cartilage growth and development. However, the mechanism by which FGF18 mediates its biological functions is still unclear. In our study, we expressed the rhFGF18 protein fused to a HaloTag, (Halo...

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Bibliographic Details
Published in:Protein expression and purification 2019-03, Vol.155, p.8-14
Main Authors: Xu, Nuo, Wang, Bao hui, Zhou, Qianyun, Ouyang, Yuehong, Gong, Weiyue, Tian, Haishan, Li, Xiaokun, Jiang, Chao
Format: Article
Language:English
Subjects:
Animals
ATDC5 cell
Cartilage - cytology
Cell Differentiation
Cell Line
Cell Proliferation
Differentiation
Fibroblast Growth Factors - genetics
Fibroblast Growth Factors - metabolism
Gene Expression
HaloTag
Humans
MAP Kinase Signaling System
Mice
NIH 3T3 Cells
Protein Binding
Recombinant Proteins - genetics
Recombinant Proteins - metabolism
rhFGF18
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Summary:Fibroblast growth factor 18 (FGF18) is a member of the fibroblast growth factor family and important in cartilage growth and development. However, the mechanism by which FGF18 mediates its biological functions is still unclear. In our study, we expressed the rhFGF18 protein fused to a HaloTag, (Halo-rhFGF18). MTT assay results indicated that both rhFGF18 and Halo-rhFGF18 have similar biological activities in NIH3T3 cells. However, basic FGF and acidic FGF were more potent than both rhFGF18 and Halo-rhFGF18. Confocal imaging data indicated that the red fluorescence labeled Halo-rhFGF18 strongly bound to ATDC5 cells and stimulated their proliferation and differentiation, which suggests that glycosaminoglycans may be involved in mediating the biological effects of rhFGF18 in ATDC5 cells. Moreover, western blot results demonstrated that, in ATDC5 cells, ERK1/2 signaling is activated upon stimulation with rhFGF18. Our results may open doors for the use of rhFGF18 as a drug to promote cartilage growth. •The halo-rhFGF18 was expressed successfully by E. coli system, purify by heparin affinity for the first time in this study•The halo-rhFGF18 improves proliferation of NIH3T3 and induces differentiation on ATDC5 more significant than non-fused FGF18.•The halo-rhFGF18 promotes chondrocyte differentiation through ERK signaling pathway were demonstrated in this study.
ISSN:1046-5928
1096-0279
DOI:10.1016/j.pep.2018.10.010