Title: BMP-4 Induction of Arrest and Differentiation of Osteoblast-Like Cells via p21(CIP1) and p27(KIP1) Regulation
Authors: Chang, Shun-Fu
Chang, Ting-Kuo
Peng, Hsin-Hsin
Yeh, Yi-Ting
Lee, Ding-Yu
Yeh, Chiuan-Ren
Zhou, Jing
Cheng, Cheng-Kung
Chang, Cheng Allen
Chiu, Jeng-Jiann
Department of Biological Science and Technology
Issue Date: 1-Nov-2009
Abstract: Cell cycle regulation by differentiation signals is critical for eukaryote development. We investigated the roles of bone morphogenetic protein (BMP)-4, an important stimulator of osteoblast differentiation and bone formation, in regulating cell cycle distribution in four osteoblast-like cell lines and mouse primary osteoblasts, and the underlying mechanisms. In all cells used, BMP-4 induced G(0)/G(1) arrest. The molecular basis of the BMP-4 effect was analyzed, and the presentation on molecular mechanism is focused on human MG63 cells. BMP-4 induced p21(CIP1) and p27(KIP1) expressions and hence cell differentiation but had no effects on the expressions of cyclins A, B1, D1, and E, cyclin-dependent protein kinase-2, -4, and -6. Using specific small interfering RNA (siRNA), we found that BMP-4-induced G(0)/G(1) arrest, and p21(CIP1) and p27(KIP1) expressions were mediated by BMP receptor type IA (BMPRIA)-specific Sma- and Mad-related protein (Smad)1/5. BMP-4 induced transient phosphorylations of ERK; transfection of MG63 cells with ERK2, but not ERK1, -specific siRNA inhibited the BMP-4-induced responses in MG63 cells. Pretreatment of MG63 cells with Arg-Gly-Asp-Ser, which blocks the cell-extracellular matrix interaction, or transfection with beta(3) integrin-specific siRNA inhibited BMP-4-induced ERK and Smad1/5 phosphorylations. BMP-4 induced transient increases in associations of beta(3)-integrin with focal adhesion kinase and Shc, the dominant-negative mutants of which inhibited BMP-4-induced ERK and Smad1/5 phosphorylations. Our results indicate that BMP-4 induces G(0)/G(1) arrest and hence differentiation in osteoblast-like cells through increased expressions of p21(CIP1) and p27(KIP1), which are mediated by BMPRIA-specific Smad1/5. The extracellular matrix/beta(3) integrin/focal adhesion kinase/Shc/ERK2 signaling pathway is involved in these BMP-4-induced responses in osteoblast-like cells. (Molecular Endocrinology 23: 1827-1838, 2009)
URI: http://dx.doi.org/10.1210/me.2009-0143
ISSN: 0888-8809
DOI: 10.1210/me.2009-0143
Volume: 23
Issue: 11
Begin Page: 1827
End Page: 1838
Appears in Collections:Articles