ECTS2014 Poster Presentations Cell biology: osteoblasts and bone formation (48 abstracts)
EGF inhibits Wnt/β-catenin-induced osteoblast differentiation in a Smurf1-dependent manner
Arang Kwon 1 , Kanitsak Boonanantanasarn 1, , Hye-Lim Lee 1 , Kyung Mi Woo 1 , Hyun-Mo Ryoo 1 , Gwan-Shik Kim 1 & Jeong-Hwa Baek 1
1Seoul National University School of Dentistry, Seoul, Republic of Korea; 2Mahidol University, Bangkok, Thailand.
BMPs and canonical Wnts are the representative developmental signals that enhance osteoblast differentiation and bone formation. Previously, we demonstrated that EGF inhibits BMP2-induced osteoblast differentiation through the induction of Smurf1 expression. However, the regulatory role of EGF in Wnt/β-catenin-induced osteoblast differentiation has not been elucidated. In this study, we investigated the effect of EGF on Wnt/β-catenin signaling-induced osteoblast differentiation using C2C12 cell line. EGF significantly suppressed the expression of osteoblast marker genes, which was induced by Wnt3a and GSK-3β inhibitor. EGF increased the expression levels of Smurf1 mRNA and protein. Smurf1 knockdown rescued Wnt/β-catenin-induced osteogenic marker gene expression in the presence of EGF. EGF treatment or Smurf1 overexpression did not affect the expression level of β-catenin mRNA but reduced β-catenin protein level and TOPFLASH activity. EGF and Smurf1 promoted β-catenin ubiquitination. Co-immunoprecipitation and GST pull-down assays showed that Smurf1 associates with β-catenin. These results suggest that EGF/Smurf1 inhibit Wnt/β-catenin-induced osteogenic differentiation and that Smurf1 downregulate Wnt/β-catenin signalling via enhancing proteasomal degradation of β-catenin.
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