Metabolism and Action of 25-Hydroxy-19-nor-Vitamin D 3 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) 2 D 3 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-D 3 -dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH) 2 -19-nor-D 3 , 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-D 3 was hardly converted to 1α,25(OH) 2 -19-nor-D 3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D 3 and 25(OH)-19-nor-D 3 demonstrated that the k cat /K m for 25(OH)-19-nor-D 3 is less than 0.1% of that for 25(OH)D 3 . When 25(OH)-19-nor-D 3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH) 2 -19-nor-D 3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D 3 , and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 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-D 3 , whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D 3 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
4 alpha-glucanotransferase
Vitamin D
Genes
Gene Knockdown Techniques
Hydroxylation
Hydroxyl Radical
Osteoporosis
Cats
Ligands
Cell Line
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 D 3 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 D 3 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 D 3 in Human Prostate Cells",
abstract = "Since the discovery of 1α,25(OH) 2 D 3 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-D 3 -dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH) 2 -19-nor-D 3 , 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-D 3 was hardly converted to 1α,25(OH) 2 -19-nor-D 3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D 3 and 25(OH)-19-nor-D 3 demonstrated that the k cat /K m for 25(OH)-19-nor-D 3 is less than 0.1{\%} of that for 25(OH)D 3 . When 25(OH)-19-nor-D 3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH) 2 -19-nor-D 3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D 3 , and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 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-D 3 , whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D 3 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 D 3 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 D 3 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 D 3 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) 2 D 3 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-D 3 -dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH) 2 -19-nor-D 3 , 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-D 3 was hardly converted to 1α,25(OH) 2 -19-nor-D 3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D 3 and 25(OH)-19-nor-D 3 demonstrated that the k cat /K m for 25(OH)-19-nor-D 3 is less than 0.1% of that for 25(OH)D 3 . When 25(OH)-19-nor-D 3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH) 2 -19-nor-D 3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D 3 , and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 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-D 3 , whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D 3 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) 2 D 3 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-D 3 -dependent growth inhibition of human prostate PZ-HPV-7 cells was the result of its subsequent conversion to 1α,25(OH) 2 -19-nor-D 3 , 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-D 3 was hardly converted to 1α,25(OH) 2 -19-nor-D 3 by the enzyme. The kinetic analysis of 1α-hydroxylation of 25(OH)D 3 and 25(OH)-19-nor-D 3 demonstrated that the k cat /K m for 25(OH)-19-nor-D 3 is less than 0.1% of that for 25(OH)D 3 . When 25(OH)-19-nor-D 3 was added to cultured PZ-HPV-7 cells, eight metabolites were detected, while no 1α,25(OH) 2 -19-nor-D 3 was found. In addition, the time course of VDR translocation into the nucleus induced by 100 nM 25(OH)-19-nor-D 3 , and the subsequent transactivation of CYP24A1 gene were almost identical to those induced by 1 nM 1α,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 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-D 3 , whereas VDR knockdown attenuated the effect, suggesting that the antiproliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but CYP27B1 independent. Finally, our recent studies using the same cell line demonstrate that 25(OH)D 3 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 D 3 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