Human prostate cells synthesize 1,25-dihydroxyvitamin D3 from 25- hydroxyvitamin D3

Gary G. Schwartz, Lyman W. Whitlatch, Tai C. Chen, Balakrishna L Lokeshwar, Michael F. Holick

Research output: Contribution to journalArticle

308 Citations (Scopus)

Abstract

Epidemiological and laboratory data support a role for vitamin D in the growth and differentiation of human prostatic cells. These findings prompted us to ask whether prostatic cells could convert 25-hydroxyvitamin D3 (25- OH-D3), the major circulating metabolite of vitamin D3, to 1,25- dihydroxyvitamin D3 [1,25(OH)2D3], the hormonally active metabolite, in a manner similar to cultured human keratinocytes. Therefore, we investigated three well-characterized human prostate cancer cell lines, LNCaP, DU 145, and PC-3; two primary cultures of cells derived from noncancerous human prostates (one normal and one benign prostatic hyperplasia); and primary cultures of normal human keratinocytes for their ability to synthesize 1,25(OH)2D3. Assays were performed in the presence of 25-OH-D3 as the enzyme substrate and 1,2-dianilinoethane, an antioxidant and free radical scavenger, and in the presence and absence of clotrimazole, a cytochrome P450 inhibitor. DU 145 and PC-3 cells produced 0.31 ± 0.06 and 0.07 ± 0.01 pmol of 1,25(OH)2D3/mg protein/h, respectively. No measurable 1,25(OH)2D3 was detected in LNCaP cells. The normal and benign prostatic hyperplasia primary cultures and keratinocyte cultures produced 3.08 ± 1.56, 1.05 ± 0.31, and 2.1 ± 0.1 pmol of 1,25(OH)2D3/mg protein/h, respectively, using a calf thymus receptor binding assay to measure 1,25(OH)2D3 in the presence of 1,2-dianilinoethane. The identity of the analyte as 1,25(OH)2D3 was supported by high performance liquid chromatography using [3H]25-OH-D3 as the enzyme substrate and a solvent system that is specific for 1,25(OH)2D3. The production of 1,25(OH)2D3 in the prostate cancer cell lines and in the primary cultures was completely inhibited in the presence of clotrimazole. This report demonstrates that two of three human prostate cancer cell lines, as well as primary cultures of noncancerous prostatic cells, possess 1α- hydroxylase activity and can synthesize 1,25(OH)2D3 from 25-OH-D3. Together with recent data indicating that 1,25(OH)2D3 inhibits the invasiveness of human prostate cancer cells (G. G. Schwartz et al., Cancer Epidemiol. Biomark. Prev., 6: 727-732, 1997), these data suggest a potential role for 25-OH-D3 in the chemoprevention of invasive prostate cancer.

Original languageEnglish (US)
Pages (from-to)391-395
Number of pages5
JournalCancer Epidemiology Biomarkers and Prevention
Volume7
Issue number5
StatePublished - May 1 1998

Fingerprint

Calcifediol
Calcitriol
Prostate
Prostatic Neoplasms
Keratinocytes
Clotrimazole
Prostatic Hyperplasia
Cell Line
Free Radical Scavengers
Primary Cell Culture
Cholecalciferol
Chemoprevention
Enzymes
Mixed Function Oxygenases
Vitamin D
Cytochrome P-450 Enzyme System
Thymus Gland
Proteins
Antioxidants
High Pressure Liquid Chromatography

ASJC Scopus subject areas

  • Epidemiology
  • Oncology

Cite this

Human prostate cells synthesize 1,25-dihydroxyvitamin D3 from 25- hydroxyvitamin D3. / Schwartz, Gary G.; Whitlatch, Lyman W.; Chen, Tai C.; Lokeshwar, Balakrishna L; Holick, Michael F.

In: Cancer Epidemiology Biomarkers and Prevention, Vol. 7, No. 5, 01.05.1998, p. 391-395.

Research output: Contribution to journalArticle

Schwartz, Gary G. ; Whitlatch, Lyman W. ; Chen, Tai C. ; Lokeshwar, Balakrishna L ; Holick, Michael F. / Human prostate cells synthesize 1,25-dihydroxyvitamin D3 from 25- hydroxyvitamin D3. In: Cancer Epidemiology Biomarkers and Prevention. 1998 ; Vol. 7, No. 5. pp. 391-395.
@article{c91581472c59468281e7e7c7a805b378,
title = "Human prostate cells synthesize 1,25-dihydroxyvitamin D3 from 25- hydroxyvitamin D3",
abstract = "Epidemiological and laboratory data support a role for vitamin D in the growth and differentiation of human prostatic cells. These findings prompted us to ask whether prostatic cells could convert 25-hydroxyvitamin D3 (25- OH-D3), the major circulating metabolite of vitamin D3, to 1,25- dihydroxyvitamin D3 [1,25(OH)2D3], the hormonally active metabolite, in a manner similar to cultured human keratinocytes. Therefore, we investigated three well-characterized human prostate cancer cell lines, LNCaP, DU 145, and PC-3; two primary cultures of cells derived from noncancerous human prostates (one normal and one benign prostatic hyperplasia); and primary cultures of normal human keratinocytes for their ability to synthesize 1,25(OH)2D3. Assays were performed in the presence of 25-OH-D3 as the enzyme substrate and 1,2-dianilinoethane, an antioxidant and free radical scavenger, and in the presence and absence of clotrimazole, a cytochrome P450 inhibitor. DU 145 and PC-3 cells produced 0.31 ± 0.06 and 0.07 ± 0.01 pmol of 1,25(OH)2D3/mg protein/h, respectively. No measurable 1,25(OH)2D3 was detected in LNCaP cells. The normal and benign prostatic hyperplasia primary cultures and keratinocyte cultures produced 3.08 ± 1.56, 1.05 ± 0.31, and 2.1 ± 0.1 pmol of 1,25(OH)2D3/mg protein/h, respectively, using a calf thymus receptor binding assay to measure 1,25(OH)2D3 in the presence of 1,2-dianilinoethane. The identity of the analyte as 1,25(OH)2D3 was supported by high performance liquid chromatography using [3H]25-OH-D3 as the enzyme substrate and a solvent system that is specific for 1,25(OH)2D3. The production of 1,25(OH)2D3 in the prostate cancer cell lines and in the primary cultures was completely inhibited in the presence of clotrimazole. This report demonstrates that two of three human prostate cancer cell lines, as well as primary cultures of noncancerous prostatic cells, possess 1α- hydroxylase activity and can synthesize 1,25(OH)2D3 from 25-OH-D3. Together with recent data indicating that 1,25(OH)2D3 inhibits the invasiveness of human prostate cancer cells (G. G. Schwartz et al., Cancer Epidemiol. Biomark. Prev., 6: 727-732, 1997), these data suggest a potential role for 25-OH-D3 in the chemoprevention of invasive prostate cancer.",
author = "Schwartz, {Gary G.} and Whitlatch, {Lyman W.} and Chen, {Tai C.} and Lokeshwar, {Balakrishna L} and Holick, {Michael F.}",
year = "1998",
month = "5",
day = "1",
language = "English (US)",
volume = "7",
pages = "391--395",
journal = "Cancer Epidemiology Biomarkers and Prevention",
issn = "1055-9965",
publisher = "American Association for Cancer Research Inc.",
number = "5",

}

TY - JOUR

T1 - Human prostate cells synthesize 1,25-dihydroxyvitamin D3 from 25- hydroxyvitamin D3

AU - Schwartz, Gary G.

AU - Whitlatch, Lyman W.

AU - Chen, Tai C.

AU - Lokeshwar, Balakrishna L

AU - Holick, Michael F.

PY - 1998/5/1

Y1 - 1998/5/1

N2 - Epidemiological and laboratory data support a role for vitamin D in the growth and differentiation of human prostatic cells. These findings prompted us to ask whether prostatic cells could convert 25-hydroxyvitamin D3 (25- OH-D3), the major circulating metabolite of vitamin D3, to 1,25- dihydroxyvitamin D3 [1,25(OH)2D3], the hormonally active metabolite, in a manner similar to cultured human keratinocytes. Therefore, we investigated three well-characterized human prostate cancer cell lines, LNCaP, DU 145, and PC-3; two primary cultures of cells derived from noncancerous human prostates (one normal and one benign prostatic hyperplasia); and primary cultures of normal human keratinocytes for their ability to synthesize 1,25(OH)2D3. Assays were performed in the presence of 25-OH-D3 as the enzyme substrate and 1,2-dianilinoethane, an antioxidant and free radical scavenger, and in the presence and absence of clotrimazole, a cytochrome P450 inhibitor. DU 145 and PC-3 cells produced 0.31 ± 0.06 and 0.07 ± 0.01 pmol of 1,25(OH)2D3/mg protein/h, respectively. No measurable 1,25(OH)2D3 was detected in LNCaP cells. The normal and benign prostatic hyperplasia primary cultures and keratinocyte cultures produced 3.08 ± 1.56, 1.05 ± 0.31, and 2.1 ± 0.1 pmol of 1,25(OH)2D3/mg protein/h, respectively, using a calf thymus receptor binding assay to measure 1,25(OH)2D3 in the presence of 1,2-dianilinoethane. The identity of the analyte as 1,25(OH)2D3 was supported by high performance liquid chromatography using [3H]25-OH-D3 as the enzyme substrate and a solvent system that is specific for 1,25(OH)2D3. The production of 1,25(OH)2D3 in the prostate cancer cell lines and in the primary cultures was completely inhibited in the presence of clotrimazole. This report demonstrates that two of three human prostate cancer cell lines, as well as primary cultures of noncancerous prostatic cells, possess 1α- hydroxylase activity and can synthesize 1,25(OH)2D3 from 25-OH-D3. Together with recent data indicating that 1,25(OH)2D3 inhibits the invasiveness of human prostate cancer cells (G. G. Schwartz et al., Cancer Epidemiol. Biomark. Prev., 6: 727-732, 1997), these data suggest a potential role for 25-OH-D3 in the chemoprevention of invasive prostate cancer.

AB - Epidemiological and laboratory data support a role for vitamin D in the growth and differentiation of human prostatic cells. These findings prompted us to ask whether prostatic cells could convert 25-hydroxyvitamin D3 (25- OH-D3), the major circulating metabolite of vitamin D3, to 1,25- dihydroxyvitamin D3 [1,25(OH)2D3], the hormonally active metabolite, in a manner similar to cultured human keratinocytes. Therefore, we investigated three well-characterized human prostate cancer cell lines, LNCaP, DU 145, and PC-3; two primary cultures of cells derived from noncancerous human prostates (one normal and one benign prostatic hyperplasia); and primary cultures of normal human keratinocytes for their ability to synthesize 1,25(OH)2D3. Assays were performed in the presence of 25-OH-D3 as the enzyme substrate and 1,2-dianilinoethane, an antioxidant and free radical scavenger, and in the presence and absence of clotrimazole, a cytochrome P450 inhibitor. DU 145 and PC-3 cells produced 0.31 ± 0.06 and 0.07 ± 0.01 pmol of 1,25(OH)2D3/mg protein/h, respectively. No measurable 1,25(OH)2D3 was detected in LNCaP cells. The normal and benign prostatic hyperplasia primary cultures and keratinocyte cultures produced 3.08 ± 1.56, 1.05 ± 0.31, and 2.1 ± 0.1 pmol of 1,25(OH)2D3/mg protein/h, respectively, using a calf thymus receptor binding assay to measure 1,25(OH)2D3 in the presence of 1,2-dianilinoethane. The identity of the analyte as 1,25(OH)2D3 was supported by high performance liquid chromatography using [3H]25-OH-D3 as the enzyme substrate and a solvent system that is specific for 1,25(OH)2D3. The production of 1,25(OH)2D3 in the prostate cancer cell lines and in the primary cultures was completely inhibited in the presence of clotrimazole. This report demonstrates that two of three human prostate cancer cell lines, as well as primary cultures of noncancerous prostatic cells, possess 1α- hydroxylase activity and can synthesize 1,25(OH)2D3 from 25-OH-D3. Together with recent data indicating that 1,25(OH)2D3 inhibits the invasiveness of human prostate cancer cells (G. G. Schwartz et al., Cancer Epidemiol. Biomark. Prev., 6: 727-732, 1997), these data suggest a potential role for 25-OH-D3 in the chemoprevention of invasive prostate cancer.

UR - http://www.scopus.com/inward/record.url?scp=0031927920&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031927920&partnerID=8YFLogxK

M3 - Article

C2 - 9610788

AN - SCOPUS:0031927920

VL - 7

SP - 391

EP - 395

JO - Cancer Epidemiology Biomarkers and Prevention

JF - Cancer Epidemiology Biomarkers and Prevention

SN - 1055-9965

IS - 5

ER -