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DOI: 10.1055/s-2006-955085
© Georg Thieme Verlag KG Stuttgart · New York
Smad3 Differently Affects Osteoblast Differentiation Depending upon its Differentiation Stage
Publication History
Received 3 April 2006
Accepted after revision 12 June 2006
Publication Date:
16 November 2006 (online)
Abstract
Smad3, a critical component of the TGF-β signaling pathways, plays an important role in the regulation of bone formation. However, how Smad3 affects osteoblast at the different differentiation stage remains still unknown. In the present study, we examined the effects of Smad3 on osteoblast phenotype by employing mouse bone marrow ST-2 cells and mouse osteoblastic MC3T3-E1 cells at the different differentiation stage. Smad3 overexpression significantly inhibited bone morphogenetic protein-2 (BMP-2)-induced ALP activity in ST-2 cells, indicating that Smad3 suppresses the commitment of pluripotent mesenchymal cells into osteoblastic cells. Smad3 increased the levels of COLI and ALP mRNA at 7 day cultures in MC3T3-E1 cells, and its effects on COL1 were decreased as the culture periods progress, although its effects on ALP were sustained during 21 day cultures. Smad3 overexpression enhanced the level of Runx2 and OCN mRNA at 14 day and 21 day cultures. Smad3 increased the levels of MGP and NPP-1 mRNA, although the extent of increase in MGP and NPP-1 was reduced and enhanced during the progression of culture period, respectively. Smad3 did not affect the level of ANK mRNA. On the other hand, Smad3 enhanced the level of MEPE mRNA at 14 and 21 day cultures, although Smad3 decreased it at 7 day cultures. In conclusion, Smad3 inhibits the osteoblastic commitment of ST-2 cells, while promotes the early stage of differentiation and maturation of osteoblastic committed MC3T3-E1 cells. Also, Smad3 enhanced the expression of mineralization-related genes at the maturation phase of MC3T3-E1 cells.
Key words
Smad3 - osteoblast - TGF-β - mesenchymal cells - mineralization
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Correspondence
Hiroshi Kaji
Division of Endocrinology/Metabolism·Neurology and Hematology/Oncology·Department of Clinical Molecular Medicine·Kobe University Graduate School of Medicine
7-5-2 Kusunoki-cho
Chuo-ku, Kobe 650-0017
Japan
Phone: +81/78/382/58 85
Fax: +81/78/382/58 99
Email: hiroshik@med.kobe-u.ac.jp