1α,25-dihydroxyvitamin D3 stimulates human SOST gene expression and sclerostin secretion
Abstract
Sclerostin, the SOST gene product, is a negative regulator of bone formation and a positive regulator of bone resorption. Here, in this study, treatment of human primary osteoblasts, including cells differentiated to an osteocyte-like stage, with 1α,25-dihydroxyvitaminD3 (1,25D) resulted in the dose-dependent increased expression of SOST mRNA. A similar effect was observed in human trabecular bone samples cultured ex vivo, and in osteocyte-like cultures of differentiated SAOS2 cells. Treatment of SAOS2 cells with 1,25D resulted in the production and secretion of sclerostin protein. In silico analysis of the human SOST gene revealed a single putative DR3-type vitamin D response element (VDRE) at position -6216 bp upstream of the transcription start site (TSS). This sequence was confirmed to have strong VDRE activity by luciferase reporter assays and electrophoretic mobility shift analysis (EMSA). Sequence substitution in the VDR/RXR half-sites abolished VDRE reporter activity and binding of nuclear proteins. A 6.3 kb fragment of the human proximal SOST promoter demonstrated responsiveness to 1,25D. The addition of the evolutionary conserved region 5 (ECR5), a known bone specific enhancer region, ahead of the 6.3 kb fragment increased basal promoter activity but did not increase 1,25D responsiveness. Site-specific mutagenesis abolished the responsiveness of the 6.3 kbmore »
- Authors:
-
- Univ. of Adelaide, SA (Australia)
- Univ. of South Australia, Adelaide, SA (Australia)
- Lawrence Livermore National Lab. (LLNL), Livermore, CA (United States). Physical and Life Sciences Directorate; Univ. of California, Merced, CA (United States)
- Publication Date:
- Research Org.:
- Lawrence Livermore National Laboratory (LLNL), Livermore, CA (United States)
- Sponsoring Org.:
- USDOE National Nuclear Security Administration (NNSA); National Health and Medical Research Council of Australia (NHMRC); National Institutes of Health (NIH)
- OSTI Identifier:
- 1891707
- Alternate Identifier(s):
- OSTI ID: 1249889
- Report Number(s):
- LLNL-JRNL-837178
Journal ID: ISSN 0303-7207; 1056922
- Grant/Contract Number:
- AC52-07NA27344; ID565372; DK075730
- Resource Type:
- Journal Article: Accepted Manuscript
- Journal Name:
- Molecular and Cellular Endocrinology
- Additional Journal Information:
- Journal Volume: 413; Journal ID: ISSN 0303-7207
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES; biological sciences; medical sciences; vitamin D; vitamin D response element; SOST; sclerostin; osteocyte; gene regulation; promoter
Citation Formats
Wijenayaka, Asiri R., Yang, Dongqing, Prideaux, Matthew, Ito, Nobuaki, Kogawa, Masakazu, Anderson, Paul H., Morris, Howard A., Solomon, Lucian B., Loots, Gabriela G., Findlay, David M., and Atkins, Gerald J. 1α,25-dihydroxyvitamin D3 stimulates human SOST gene expression and sclerostin secretion. United States: N. p., 2015.
Web. doi:10.1016/j.mce.2015.06.021.
Wijenayaka, Asiri R., Yang, Dongqing, Prideaux, Matthew, Ito, Nobuaki, Kogawa, Masakazu, Anderson, Paul H., Morris, Howard A., Solomon, Lucian B., Loots, Gabriela G., Findlay, David M., & Atkins, Gerald J. 1α,25-dihydroxyvitamin D3 stimulates human SOST gene expression and sclerostin secretion. United States. https://doi.org/10.1016/j.mce.2015.06.021
Wijenayaka, Asiri R., Yang, Dongqing, Prideaux, Matthew, Ito, Nobuaki, Kogawa, Masakazu, Anderson, Paul H., Morris, Howard A., Solomon, Lucian B., Loots, Gabriela G., Findlay, David M., and Atkins, Gerald J. 2015.
"1α,25-dihydroxyvitamin D3 stimulates human SOST gene expression and sclerostin secretion". United States. https://doi.org/10.1016/j.mce.2015.06.021. https://www.osti.gov/servlets/purl/1891707.
@article{osti_1891707,
title = {1α,25-dihydroxyvitamin D3 stimulates human SOST gene expression and sclerostin secretion},
author = {Wijenayaka, Asiri R. and Yang, Dongqing and Prideaux, Matthew and Ito, Nobuaki and Kogawa, Masakazu and Anderson, Paul H. and Morris, Howard A. and Solomon, Lucian B. and Loots, Gabriela G. and Findlay, David M. and Atkins, Gerald J.},
abstractNote = {Sclerostin, the SOST gene product, is a negative regulator of bone formation and a positive regulator of bone resorption. Here, in this study, treatment of human primary osteoblasts, including cells differentiated to an osteocyte-like stage, with 1α,25-dihydroxyvitaminD3 (1,25D) resulted in the dose-dependent increased expression of SOST mRNA. A similar effect was observed in human trabecular bone samples cultured ex vivo, and in osteocyte-like cultures of differentiated SAOS2 cells. Treatment of SAOS2 cells with 1,25D resulted in the production and secretion of sclerostin protein. In silico analysis of the human SOST gene revealed a single putative DR3-type vitamin D response element (VDRE) at position -6216 bp upstream of the transcription start site (TSS). This sequence was confirmed to have strong VDRE activity by luciferase reporter assays and electrophoretic mobility shift analysis (EMSA). Sequence substitution in the VDR/RXR half-sites abolished VDRE reporter activity and binding of nuclear proteins. A 6.3 kb fragment of the human proximal SOST promoter demonstrated responsiveness to 1,25D. The addition of the evolutionary conserved region 5 (ECR5), a known bone specific enhancer region, ahead of the 6.3 kb fragment increased basal promoter activity but did not increase 1,25D responsiveness. Site-specific mutagenesis abolished the responsiveness of the 6.3 kb promoter to 1,25D. We conclude that 1,25D is a direct regulator of human SOST gene and sclerostin protein expression, extending the pathways of control of sclerostin expression. At least some of this responsiveness is mediated by the identified classical VDRE however the nature of the transcriptional regulation by 1,25D warrants further investigation.},
doi = {10.1016/j.mce.2015.06.021},
url = {https://www.osti.gov/biblio/1891707},
journal = {Molecular and Cellular Endocrinology},
issn = {0303-7207},
number = ,
volume = 413,
place = {United States},
year = {Tue Jun 23 00:00:00 EDT 2015},
month = {Tue Jun 23 00:00:00 EDT 2015}
}
Web of Science
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