Mechanism of the anti-proliferative action of 25-hydroxy-19-nor-vitamin D 3 in human prostate cells

Eiji Munetsuna, Sachie Nakabayashi, Rie Kawanami, Kaori Yasuda, Miho Ohta, Midori A. Arai, Atsushi Kittaka, Tai C. Chen, Masaki Kamakura, Shinichi Ikushiro, Toshiyuki Sakaki

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Abstract

According to the prevailing paradigm, 1α-hydroxylation of 25-hydroxyvitamin D 3 (25(OH)D 3 ) and its analogs is a prerequisite step for their biological effects. We previously reported that 25-hydroxy-19-nor-vitamin D 3 (25(OH)-19-nor-D 3 ) had anti-proliferative activity in a cell line, PZ-HPV-7, which was derived from human non-cancerous prostate tissue, and suggested that 25(OH)-19-nor-D 3 acted after 1α-hydroxylation by vitamin D 1α-hydroxylase (CYP27B1). However, metabolic studies of 25(OH)-19-nor-D 3 using recombinant CYP27B1 revealed that 25(OH)-19-nor-D 3 was rarely subjected to 1a-hydroxylation. Therefore, in this report, we attempted to clarify the mechanism of 25(OH)-19-nor-D 3 action in intact cells using PZ-HPV-7 prostate cells. After incubating the cells with 25(OH)-19-nor-D 3 , eight metabolites of 24-hydroxylase (CYP24A1) were detected, whereas no products of CYP27B1 including 1α,25-dihydroxy-19-nor-vitamin D 3 (1α,25(OH) 2 -19-nor-D 3 ) were found. Furthermore, the time-dependent nuclear translocation of vitamin D receptor (VDR) and the subsequent transactivation of cyp24A1 gene in the presence of 25(OH)-19-nor-D 3 were almost identical as those induced by 1a,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 directly binds to VDR as a ligand and transports VDR into the nucleus to induce transcription of cyp24A1 gene. In addition, knock down of cyp27B1 gene did not affect the anti-proliferative activity of 25(OH)-19-nor-D 3 , whereas knock down of VDR attenuated the inhibitory effect. Thus, our results clearly demonstrate that the anti-proliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but 1α-hydroxylation independent, suggesting that 25(OH)D 3 analogs such as 25(OH)-19-nor-D 3 could be attractive candidates for anticancer therapy.

Original languageEnglish
Pages (from-to)209-218
Number of pages10
JournalJournal of Molecular Endocrinology
Volume47
Issue number2
DOIs
Publication statusPublished - 01-10-2011

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Calcitriol Receptors
Cholecalciferol
25-Hydroxyvitamin D3 1-alpha-Hydroxylase
Prostate
Hydroxylation
Mixed Function Oxygenases
Gene Knockdown Techniques
Calcifediol
Vitamin D
Transcriptional Activation
Genes
Ligands
Cell Line

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Endocrinology

Cite this

Munetsuna, Eiji ; Nakabayashi, Sachie ; Kawanami, Rie ; Yasuda, Kaori ; Ohta, Miho ; Arai, Midori A. ; Kittaka, Atsushi ; Chen, Tai C. ; Kamakura, Masaki ; Ikushiro, Shinichi ; Sakaki, Toshiyuki. / Mechanism of the anti-proliferative action of 25-hydroxy-19-nor-vitamin D 3 in human prostate cells In: Journal of Molecular Endocrinology. 2011 ; Vol. 47, No. 2. pp. 209-218.
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author = "Eiji Munetsuna and Sachie Nakabayashi and Rie Kawanami and Kaori Yasuda and Miho Ohta and Arai, {Midori A.} and Atsushi Kittaka and Chen, {Tai C.} and Masaki Kamakura and Shinichi Ikushiro and Toshiyuki Sakaki",
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Munetsuna, E, Nakabayashi, S, Kawanami, R, Yasuda, K, Ohta, M, Arai, MA, Kittaka, A, Chen, TC, Kamakura, M, Ikushiro, S & Sakaki, T 2011, ' Mechanism of the anti-proliferative action of 25-hydroxy-19-nor-vitamin D 3 in human prostate cells ', Journal of Molecular Endocrinology, vol. 47, no. 2, pp. 209-218. https://doi.org/10.1530/JME-11-0008

Mechanism of the anti-proliferative action of 25-hydroxy-19-nor-vitamin D 3 in human prostate cells . / Munetsuna, Eiji; Nakabayashi, Sachie; Kawanami, Rie; Yasuda, Kaori; Ohta, Miho; Arai, Midori A.; Kittaka, Atsushi; Chen, Tai C.; Kamakura, Masaki; Ikushiro, Shinichi; Sakaki, Toshiyuki.

In: Journal of Molecular Endocrinology, Vol. 47, No. 2, 01.10.2011, p. 209-218.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Mechanism of the anti-proliferative action of 25-hydroxy-19-nor-vitamin D 3 in human prostate cells

AU - Munetsuna, Eiji

AU - Nakabayashi, Sachie

AU - Kawanami, Rie

AU - Yasuda, Kaori

AU - Ohta, Miho

AU - Arai, Midori A.

AU - Kittaka, Atsushi

AU - Chen, Tai C.

AU - Kamakura, Masaki

AU - Ikushiro, Shinichi

AU - Sakaki, Toshiyuki

PY - 2011/10/1

Y1 - 2011/10/1

N2 - According to the prevailing paradigm, 1α-hydroxylation of 25-hydroxyvitamin D 3 (25(OH)D 3 ) and its analogs is a prerequisite step for their biological effects. We previously reported that 25-hydroxy-19-nor-vitamin D 3 (25(OH)-19-nor-D 3 ) had anti-proliferative activity in a cell line, PZ-HPV-7, which was derived from human non-cancerous prostate tissue, and suggested that 25(OH)-19-nor-D 3 acted after 1α-hydroxylation by vitamin D 1α-hydroxylase (CYP27B1). However, metabolic studies of 25(OH)-19-nor-D 3 using recombinant CYP27B1 revealed that 25(OH)-19-nor-D 3 was rarely subjected to 1a-hydroxylation. Therefore, in this report, we attempted to clarify the mechanism of 25(OH)-19-nor-D 3 action in intact cells using PZ-HPV-7 prostate cells. After incubating the cells with 25(OH)-19-nor-D 3 , eight metabolites of 24-hydroxylase (CYP24A1) were detected, whereas no products of CYP27B1 including 1α,25-dihydroxy-19-nor-vitamin D 3 (1α,25(OH) 2 -19-nor-D 3 ) were found. Furthermore, the time-dependent nuclear translocation of vitamin D receptor (VDR) and the subsequent transactivation of cyp24A1 gene in the presence of 25(OH)-19-nor-D 3 were almost identical as those induced by 1a,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 directly binds to VDR as a ligand and transports VDR into the nucleus to induce transcription of cyp24A1 gene. In addition, knock down of cyp27B1 gene did not affect the anti-proliferative activity of 25(OH)-19-nor-D 3 , whereas knock down of VDR attenuated the inhibitory effect. Thus, our results clearly demonstrate that the anti-proliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but 1α-hydroxylation independent, suggesting that 25(OH)D 3 analogs such as 25(OH)-19-nor-D 3 could be attractive candidates for anticancer therapy.

AB - According to the prevailing paradigm, 1α-hydroxylation of 25-hydroxyvitamin D 3 (25(OH)D 3 ) and its analogs is a prerequisite step for their biological effects. We previously reported that 25-hydroxy-19-nor-vitamin D 3 (25(OH)-19-nor-D 3 ) had anti-proliferative activity in a cell line, PZ-HPV-7, which was derived from human non-cancerous prostate tissue, and suggested that 25(OH)-19-nor-D 3 acted after 1α-hydroxylation by vitamin D 1α-hydroxylase (CYP27B1). However, metabolic studies of 25(OH)-19-nor-D 3 using recombinant CYP27B1 revealed that 25(OH)-19-nor-D 3 was rarely subjected to 1a-hydroxylation. Therefore, in this report, we attempted to clarify the mechanism of 25(OH)-19-nor-D 3 action in intact cells using PZ-HPV-7 prostate cells. After incubating the cells with 25(OH)-19-nor-D 3 , eight metabolites of 24-hydroxylase (CYP24A1) were detected, whereas no products of CYP27B1 including 1α,25-dihydroxy-19-nor-vitamin D 3 (1α,25(OH) 2 -19-nor-D 3 ) were found. Furthermore, the time-dependent nuclear translocation of vitamin D receptor (VDR) and the subsequent transactivation of cyp24A1 gene in the presence of 25(OH)-19-nor-D 3 were almost identical as those induced by 1a,25(OH) 2 -19-nor-D 3 . These results strongly suggest that 25(OH)-19-nor-D 3 directly binds to VDR as a ligand and transports VDR into the nucleus to induce transcription of cyp24A1 gene. In addition, knock down of cyp27B1 gene did not affect the anti-proliferative activity of 25(OH)-19-nor-D 3 , whereas knock down of VDR attenuated the inhibitory effect. Thus, our results clearly demonstrate that the anti-proliferative activity of 25(OH)-19-nor-D 3 is VDR dependent but 1α-hydroxylation independent, suggesting that 25(OH)D 3 analogs such as 25(OH)-19-nor-D 3 could be attractive candidates for anticancer therapy.

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