Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells

Eiji Munetsuna, Atsushi Kittaka, Tai C. Chen, Toshiyuki Sakaki

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

Since the discovery of 1α,25(OH)2D3 in the early 1970s, it has been widely accepted that this metabolite is responsible for the biological actions of vitamin D. Likewise, we have assumed that 25(OH)-19-nor-D3-dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH)2-19-nor-D3, catalyzed by CYP27B1 within the prostate cells. However, further in vitro studies in a reconstituted system using recombinant CYP27B1 revealed that 25(OH)-19-nor-D3 was hardly converted to 1α,25(OH)2-19-nor-D3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D3 and 25(OH)-19-nor-D3 demonstrated that the kcat/Km for 25(OH)-19-nor-D3 is less than 0.1% of that for 25(OH)D3. When 25(OH)-19-nor-D3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH)2-19-nor-D3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D3, and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH)2-19-nor-D3. These results strongly suggest that 25(OH)-19-nor-D3 binds directly to VDR as a ligand to transport VDR into the nucleus to induce CYP24A1 gene transactivation. Furthermore, knockdown of CYP27B1 gene did not affect the antiproliferative activity of 25(OH)-19-nor-D3, whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D3 can act as a VDR agonist to induce gene transactivation. These findings suggest that vitamin D analogs without 1α-hydroxyl group could be developed as drugs for osteoporosis or cancer treatment.

Original languageEnglish
Title of host publicationVitamin D Hormone, 2016
EditorsGerald Litwack
PublisherAcademic Press Inc.
Pages357-377
Number of pages21
ISBN (Print)9780128048245
DOIs
Publication statusPublished - 01-01-2016

Publication series

NameVitamins and Hormones
Volume100
ISSN (Print)0083-6729

Fingerprint

25-Hydroxyvitamin D3 1-alpha-Hydroxylase
Cholecalciferol
Prostate
Transcriptional Activation
Vitamin D
Genes
Gene Knockdown Techniques
Hydroxylation
Hydroxyl Radical
Osteoporosis
Ligands
Cell Line
Enzymes
Growth
Pharmaceutical Preparations
Neoplasms

All Science Journal Classification (ASJC) codes

  • Physiology
  • Endocrinology

Cite this

Munetsuna, E., Kittaka, A., Chen, T. C., & Sakaki, T. (2016). Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells. In G. Litwack (Ed.), Vitamin D Hormone, 2016 (pp. 357-377). (Vitamins and Hormones; Vol. 100). Academic Press Inc.. https://doi.org/10.1016/bs.vh.2015.10.009
Munetsuna, Eiji ; Kittaka, Atsushi ; Chen, Tai C. ; Sakaki, Toshiyuki. / Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells. Vitamin D Hormone, 2016. editor / Gerald Litwack. Academic Press Inc., 2016. pp. 357-377 (Vitamins and Hormones).
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title = "Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells",
abstract = "Since the discovery of 1α,25(OH)2D3 in the early 1970s, it has been widely accepted that this metabolite is responsible for the biological actions of vitamin D. Likewise, we have assumed that 25(OH)-19-nor-D3-dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH)2-19-nor-D3, catalyzed by CYP27B1 within the prostate cells. However, further in vitro studies in a reconstituted system using recombinant CYP27B1 revealed that 25(OH)-19-nor-D3 was hardly converted to 1α,25(OH)2-19-nor-D3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D3 and 25(OH)-19-nor-D3 demonstrated that the kcat/Km for 25(OH)-19-nor-D3 is less than 0.1{\%} of that for 25(OH)D3. When 25(OH)-19-nor-D3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH)2-19-nor-D3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D3, and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH)2-19-nor-D3. These results strongly suggest that 25(OH)-19-nor-D3 binds directly to VDR as a ligand to transport VDR into the nucleus to induce CYP24A1 gene transactivation. Furthermore, knockdown of CYP27B1 gene did not affect the antiproliferative activity of 25(OH)-19-nor-D3, whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D3 can act as a VDR agonist to induce gene transactivation. These findings suggest that vitamin D analogs without 1α-hydroxyl group could be developed as drugs for osteoporosis or cancer treatment.",
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Munetsuna, E, Kittaka, A, Chen, TC & Sakaki, T 2016, Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells. in G Litwack (ed.), Vitamin D Hormone, 2016. Vitamins and Hormones, vol. 100, Academic Press Inc., pp. 357-377. https://doi.org/10.1016/bs.vh.2015.10.009

Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells. / Munetsuna, Eiji; Kittaka, Atsushi; Chen, Tai C.; Sakaki, Toshiyuki.

Vitamin D Hormone, 2016. ed. / Gerald Litwack. Academic Press Inc., 2016. p. 357-377 (Vitamins and Hormones; Vol. 100).

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells

AU - Munetsuna, Eiji

AU - Kittaka, Atsushi

AU - Chen, Tai C.

AU - Sakaki, Toshiyuki

PY - 2016/1/1

Y1 - 2016/1/1

N2 - Since the discovery of 1α,25(OH)2D3 in the early 1970s, it has been widely accepted that this metabolite is responsible for the biological actions of vitamin D. Likewise, we have assumed that 25(OH)-19-nor-D3-dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH)2-19-nor-D3, catalyzed by CYP27B1 within the prostate cells. However, further in vitro studies in a reconstituted system using recombinant CYP27B1 revealed that 25(OH)-19-nor-D3 was hardly converted to 1α,25(OH)2-19-nor-D3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D3 and 25(OH)-19-nor-D3 demonstrated that the kcat/Km for 25(OH)-19-nor-D3 is less than 0.1% of that for 25(OH)D3. When 25(OH)-19-nor-D3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH)2-19-nor-D3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D3, and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH)2-19-nor-D3. These results strongly suggest that 25(OH)-19-nor-D3 binds directly to VDR as a ligand to transport VDR into the nucleus to induce CYP24A1 gene transactivation. Furthermore, knockdown of CYP27B1 gene did not affect the antiproliferative activity of 25(OH)-19-nor-D3, whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D3 can act as a VDR agonist to induce gene transactivation. These findings suggest that vitamin D analogs without 1α-hydroxyl group could be developed as drugs for osteoporosis or cancer treatment.

AB - Since the discovery of 1α,25(OH)2D3 in the early 1970s, it has been widely accepted that this metabolite is responsible for the biological actions of vitamin D. Likewise, we have assumed that 25(OH)-19-nor-D3-dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH)2-19-nor-D3, catalyzed by CYP27B1 within the prostate cells. However, further in vitro studies in a reconstituted system using recombinant CYP27B1 revealed that 25(OH)-19-nor-D3 was hardly converted to 1α,25(OH)2-19-nor-D3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D3 and 25(OH)-19-nor-D3 demonstrated that the kcat/Km for 25(OH)-19-nor-D3 is less than 0.1% of that for 25(OH)D3. When 25(OH)-19-nor-D3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH)2-19-nor-D3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D3, and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH)2-19-nor-D3. These results strongly suggest that 25(OH)-19-nor-D3 binds directly to VDR as a ligand to transport VDR into the nucleus to induce CYP24A1 gene transactivation. Furthermore, knockdown of CYP27B1 gene did not affect the antiproliferative activity of 25(OH)-19-nor-D3, whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D3 can act as a VDR agonist to induce gene transactivation. These findings suggest that vitamin D analogs without 1α-hydroxyl group could be developed as drugs for osteoporosis or cancer treatment.

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Munetsuna E, Kittaka A, Chen TC, Sakaki T. Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D3 in Human Prostate Cells. In Litwack G, editor, Vitamin D Hormone, 2016. Academic Press Inc. 2016. p. 357-377. (Vitamins and Hormones). https://doi.org/10.1016/bs.vh.2015.10.009