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Type: Journal article
Title: RNAi-mediated silencing of CYP27B1 abolishes 1,25(OH)₂D₃ synthesis and reduces osteocalcin and CYP24 mRNA expression in human osteosarcoma (HOS) cells
Other Titles: RNAi-mediated silencing of CYP27B1 abolishes 1,25(OH)(2)D(3) synthesis and reduces osteocalcin and CYP24 mRNA expression in human osteosarcoma (HOS) cells
Author: Anderson, P.
Atkins, G.
Findlay, D.
O'Loughlin, P.
Welldon, K.
Vincent, C.
Morris, H.
Citation: Journal of Steroid Biochemistry and Molecular Biology, 2007; 103(3-5 Sp Iss):601-605
Publisher: Pergamon-Elsevier Science Ltd
Issue Date: 2007
ISSN: 0960-0760
Statement of
P.H. Anderson, G.J. Atkins, D.M. Findlay, P.D. Oloughlin, K. Welldon, C. Vincent and H.A. Morris
Abstract: Although local synthesis of 1,25D has been postulated to regulate parameters of cell growth and differentiation in non-renal cells, the physiological role of 1,25D production in bone cells remains unclear. We used the technique of RNA interference to inhibit the mRNA encoding the enzyme responsible for 1,25D synthesis, 25-hydroxyvitamin D 1α-hydroxylase (CYP27B1). Human osteosarcoma (HOS) cells were transfected with siRNA for CYP27B1 or non-silencing RNA before being treated with 25D for 48 h under normal growth conditions. De novo synthesis of 1,25D was measured in the media as well as mRNA levels for CYP27B1, osteocalcin (OCN) and 25-hydroxyvitamin D 24-hydroxylase (CYP24). We demonstrated that HOS cells express CYP27B1 mRNA, metabolize 25D and secrete detectable levels of de novo synthesized 1,25D. CYP27B1 mRNA silencing by RNAi, resulted in the suppression of 1,25D production and subsequent reduction of OCN and CYP24 mRNA expression. Our findings suggest that local 1,25D synthesis has paracrine effects in the bone microenvironment implying that vitamin D metabolism in human osteoblasts represents a physiologically important pathway, possibly regulating the maturation of osteoblasts.
Keywords: Osteoblast; Osteocalcin; 25-Hydroxyvitamin D 1α-hydroxylase; 25-Hydroxyvitamin D 24-hydroxylase; RNAi
Description: Special Issue: Proceedings of the 13th Workshop on Vitamin D (Victoria, British Columbia, Canada, April 2006). Edited by R. Bouillon, A.W. Norman and J.R. Pasqualini.
Rights: Crown copyright © 2006 Published by Elsevier Ltd.
RMID: 0020070689
DOI: 10.1016/j.jsbmb.2006.12.084
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Appears in Collections:Molecular and Biomedical Science publications

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