Core-binding factor α1 (Cbfa1) induces osteoblastic differentiation of C2C12 cells without interactions with Smad1 and Smad5

Nishimura, Riko; Hata, Kenji; Harris, S. E.; Ikeda, Fumiyo; Yoneda, Toshiyuki

doi:10.1016/s8756-3282(02)00826-8

Cited by 125 publications

(113 citation statements)

References 28 publications

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“…In one previous study, Runx2 was ectopic, expressed in C2C12 cells that were cotransduced with Smad6, an antagonist of Smad1/5 signaling, and the Runx2 overexpression still sufficiently induced osteoblastic differentiation in these cells. (44) These data indicate that the regulation of Runx2 transcriptional activity is by Smadindependent pathways. Moreover, in our study, Runx2 transcriptional activity and overexpression of Runx2-induced osteoblastic differentiation are enhanced by JNK1 inhibition.…”

Section: Discussionmentioning

confidence: 82%

c-Jun N-terminal kinase 1 negatively regulates osteoblastic differentiation induced by BMP2 via phosphorylation of Runx2 at Ser104

Huang¹,

Lin²,

Chao³

et al. 2012

Journal of Bone and Mineral Research

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Runx2 plays a crucial role in osteoblastic differentiation, which can be upregulated by bone morphogenetic proteins 2 (BMP2). Mitogenactivated protein kinase (MAPK) cascades, such as extracellular signal-regulated kinase (ERK) and p38, have been reported to be activated by BMP2 to increase Runx2 activity. The role of cjun-N-terminal kinase (JNK), the other kinase of MAPK, in osteoblastic differentiation has not been well elucidated. In this study, we first showed that JNK1 is activated by BMP2 in multipotent C2C12 and preosteoblastic MC3T3-E1 cell lines. We then showed that early and late osteoblastic differentiation, represented by ALP expression and mineralization, respectively, are significantly enhanced by JNK1 loss-of-function, such as treatment of JNK inhibitor, knockdown of JNK1 and ectopic expression of a dominant negative JNK1 (DN-JNK1). Consistently, BMP2-induced osteoblastic differentiation is reduced by JNK1 gain-offunction, such as enforced expression of a constitutively active JNK1 (CA-JNK1). Most importantly, we showed that Runx2 is required for JNK1-mediated inhibition of osteoblastic differentiation, and identified Ser104 of Runx2 is the site phosphorylated by JNK1 upon BMP2 stimulation. Finally, we found that overexpression of the mutant Runx2 (Ser104Ala) stimulates osteoblastic differentiation of C2C12 and MC3T3-E1 cells to the extent similar to that achieved by overexpression of wild-type (WT) Runx2 plus JNK inhibitor treatment. Taken together, these data indicate that JNK1 negatively regulates BMP2-induced osteoblastic differentiation through phosphorylation of Runx2 at Ser104. In addition, unraveling these mechanisms may help to develop new strategies in enhancing osteoblastic differentiation and bone formation. ß

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Section: Discussionmentioning

confidence: 82%

c-Jun N-terminal kinase 1 negatively regulates osteoblastic differentiation induced by BMP2 via phosphorylation of Runx2 at Ser104

Huang¹,

Lin²,

Chao³

et al. 2012

Journal of Bone and Mineral Research

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“…Adenoviruses carrying Runx2, Smad6, or Noggin were used as described previously (Nishimura et al, 2002;Nifuji et al, 2004). The viruses showed no proliferative activity due to a lack of E1A-E1B (Nishimura et al, 2002). Titers of the viruses were determined using a modified point assay (Nishimura et al, 2002).…”

Section: Generation Of Adenovirusmentioning

confidence: 99%

“…The viruses showed no proliferative activity due to a lack of E1A-E1B (Nishimura et al, 2002). Titers of the viruses were determined using a modified point assay (Nishimura et al, 2002). Infection of recombinant adenoviruses with C3H10T1/2 cells, C2C12 cells, or primary osteoblasts was performed by incubation with the adenoviruses at a multiplicity of infection (moi) of 40, except where specifically indicated.…”

Section: Generation Of Adenovirusmentioning

confidence: 99%

“…The cells were washed four times with ice-cold phosphate-buffered saline (PBS) and solubilized in lysis buffer (Nishimura et al, 2002). The lysates were then centrifuged for 20 min at 4°C at 16,000 ϫ g and incubated with antibodies for 4 h at 4°C, followed by immunoprecipitation with protein A-Sepharose (Zymed, South San Francisco, CA) or protein G-agarose (Roche).…”

Section: Immunoprecipitation and Immunoblottingmentioning

confidence: 99%

“…Consistently, Ihh and BMP2 synergistically stimulate osteoblast differentiation of mesenchymal cells (Nakamura et al, 1997), suggesting the cooperative role of Ihh and BMP2 signaling in osteoblast development. Runx2 (Cbfa1/ PEBP2␣A/Osf2), an essential transcription factor for bone formation and osteoblast differentiation, has been shown to function as a downstream of BMP2 signaling during osteoblastogenesis (Zhang et al, 2000;Nishimura et al, 2002). Therefore, it is possible that Ihh regulates Runx2 during osteoblast differentiation of mesenchymal cells.…”

Section: Introductionmentioning

confidence: 99%

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Ihh/Gli2 Signaling Promotes Osteoblast Differentiation by Regulating Runx2 Expression and Function

Shimoyama

Wada

Ikeda

et al. 2007

MBoC

155

115

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Genetic and cell biological studies have indicated that Indian hedgehog (Ihh) plays an important role in bone development and osteoblast differentiation. However, the molecular mechanism by which Ihh regulates osteoblast differentiation is complex and remains to be fully elucidated. In this study, we investigated the role of Ihh signaling in osteoblast differentiation using mesenchymal cells and primary osteoblasts. We observed that Ihh stimulated alkaline phosphatase (ALP) activity, osteocalcin expression, and calcification. Overexpression of Gli2-but not Gli3-induced ALP, osteocalcin expression, and calcification of these cells. In contrast, dominant-negative Gli2 markedly inhibited Ihh-dependent osteoblast differentiation. Ihh treatment or Gli2 overexpression also up-regulated the expression of Runx2, an essential transcription factor for osteoblastogenesis, and enhanced the transcriptional activity and osteogenic action of Runx2. Coimmunoprecipitation analysis demonstrated a physical interaction between Gli2 and Runx2. Moreover, Ihh or Gli2 overexpression failed to increase ALP activity in Runx2-deficient mesenchymal cells. Collectively, these results suggest that Ihh regulates osteoblast differentiation of mesenchymal cells through up-regulation of the expression and function of Runx2 by Gli2.

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Synthesis and self‐assembling behaviors of biotinylated pluronic/poly(lactic acid) biocompatible block copolymers in aqueous solutions

Li¹,

Xiong²,

Li³

et al. 2009

J of Applied Polymer Sci

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Four kinds of Biotinylated Pluronic/PLA block copolymers were synthesized by two-step reactions. Pluronic were firstly modified by biotin to obtain B-Pluronic-OH. Biotin-Pluronic-PLA block copolymers were then produced by ring-opening polymerization of the monomer L-lactide using Biotin-Pluronic-OH as the initiator and stannous octoate (Sn(Oct) 2 ) as the catalyst. The self-assembling behaviors of Biotin-Pluronic-PLA block copolymers in aqueous solutions were examined by fluorescence measurement, dynamic light scattering (DLS), and transmission electron microscopic (TEM) techniques. The size of Biotin-F127-PLA-61, Biotin-F87-PLA, and Biotin-P85-PLA nanoparticles were determined to be 198, 229, and 257 nm, respectively, and their morphologies were found to be spherical micelles. Biotin-F127-PLA-87 produces both spherical micelles and large compound micelles with the size of 127 and 906 nm. The cytotoxicity studies using human ovarian cancer cells OVCAR-3 indicate that Biotin-Pluronic-PLA block copolymers have good biocompatibility. V C 2009 Wiley Periodicals, Inc. J Appl Polym

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Core-binding factor α1 (Cbfa1) induces osteoblastic differentiation of C2C12 cells without interactions with Smad1 and Smad5

Cited by 125 publications

References 28 publications

c-Jun N-terminal kinase 1 negatively regulates osteoblastic differentiation induced by BMP2 via phosphorylation of Runx2 at Ser104

c-Jun N-terminal kinase 1 negatively regulates osteoblastic differentiation induced by BMP2 via phosphorylation of Runx2 at Ser104

Ihh/Gli2 Signaling Promotes Osteoblast Differentiation by Regulating Runx2 Expression and Function

Synthesis and self‐assembling behaviors of biotinylated pluronic/poly(lactic acid) biocompatible block copolymers in aqueous solutions

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