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Osterix is Required for Sonic Hedgehog-Induced Osteoblastic MC3T3-E1 Cell Differentiation

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Abstract

It has been shown that hedgehog (Hh) signaling plays an important role during bone development. However, the mechanism(s) by which Hh stimulates osteoblast differentiation are not fully elucidated. This study was performed to examine if Sonic hedgehog (Shh) affects osteoblast differentiation in osteoblastic MC3T3-E1 cells and determine the exact role of osterix (Osx) involved in Hh-induced osteoblast differentiation. Our real-time RT-PCR result shows that Shh significantly induced osteoblast differentiation by up-regulation of osteocalcin, alkaline phosphatase, bone sialoprotein, Type I collagen, runt-related transcription factor 2 (Runx2), and Osx RNA expression in MC3T3-E1 cells. ALP protein activity, Osx protein expression, as well as Osx promoter activity was also enhanced by Shh treatment in cell culture. Interestingly, Shh-induced Osx up-regulation was only slightly affected by knocking down Runx2 using siRNA in cells within 3 days of culture, however, knocking down Osx expression in cells totally blocked Shh-induced osteoblast differentiation. These findings demonstrate for the first time that Shh stimulates early osteoblast differentiation mainly through up-regulation of the expression of Osx in osteoblastic MC3T3-E1 cells in both Runx2-dependent and Runx2-independent manner.

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Acknowledgments

The authors are grateful to the Research Grant awarded by Ministry of Science and Technology in Shenyang City, China for partial funding support. We would like to thank Central Laboratory, Shengjing Hospital of China Medical University for providing the necessary facilities and equipments to carry out this research.

Conflict of interest

All authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Orthopaedics, Shengjing Hospital of China Medical University, Sanhao Street 36, Heping District, Shenyang, 110004, China

    Ye Tian & Qin Fu

  2. Department of Anesthesiology, Shengjing Hospital of China Medical University, Sanhao Street 36, Heping District, Shenyang, 110004, China

    Ying Xu

  3. Center for Musculoskeletal Research, University of Rochester Medical Center, Rochester, NY, 14642, USA

    Yufeng Dong

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  1. Ye Tian
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  2. Ying Xu
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Correspondence to Ye Tian.

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Tian, Y., Xu, Y., Fu, Q. et al. Osterix is Required for Sonic Hedgehog-Induced Osteoblastic MC3T3-E1 Cell Differentiation. Cell Biochem Biophys 64, 169–176 (2012). https://doi.org/10.1007/s12013-012-9369-7

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  • DOI: https://doi.org/10.1007/s12013-012-9369-7

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