Regulation of androgen and vitamin D receptors by 1,25-dihydroxyvitamin D3 in human prostate epithelial and stromal cells

Eddy S. Leman, Fernando DeMiguel, Allen C Gao, Robert H. Getzenberg

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Purpose: The mechanisms of the interaction between 1,25-dihydroxyvitamin D3 (1,25 D) and androgens, and their respective receptors in their action on the prostate are not completely understood. We examined the interplay of 1,25 D and androgens on the epithelial and stromal cells of the prostate. Materials and Methods: The human neonatal prostatic epithelial cell line 267B-1 (BRFF, Inc., Ijamsville, Maryland) and primary cultures of human prostate stromal cells were treated with medium containing 5 or 10 μM 1,25 D or ethanol (control) in the presence or absence of 10 nM dihydrotestosterone (DHT) (Sigma Chemical Co., St. Louis, Missouri). Protein levels of androgen receptor (AR) and vitamin D receptor (VDR) were determined by immunoblot analysis of whole cell extracts. Electrophoresis mobility shift assays were used to determine AR and VDR DNA binding activities. Results: The VDR protein level of 267B-1 cells was increased in the presence of 1,25 D (with the maximum effects seen at 24 hours) regardless of the presence or absence of DHT. In addition, exogenous DHT increased the AR and VDR DNA binding activities of 267B-1 and stromal cells in the presence of 1,25 D. Conclusions: ARs in the normal prostate are regulated by androgens, whereas VDRs in the normal prostate can be regulated by 1,25 D as well as by other androgens such as testosterone. This finding further supports the concept that 1,25 D as a steroid hormone, in addition to other androgens such as DHT, may have a role in the growth and differentiation of normal prostate.

Original languageEnglish (US)
Pages (from-to)235-240
Number of pages6
JournalJournal of Urology
Volume170
Issue number1
DOIs
StatePublished - Jul 1 2003
Externally publishedYes

Fingerprint

Calcitriol Receptors
Calcitriol
Stromal Cells
Androgens
Prostate
Dihydrotestosterone
Epithelial Cells
Androgen Receptors
DNA
Electrophoretic Mobility Shift Assay
Cell Extracts
Electrophoresis
Testosterone
Proteins
Ethanol
Steroids
Hormones
Cell Line
Growth

Keywords

  • Androgens
  • Calcitriol
  • Prostate
  • Receptors, androgens
  • Steroids

ASJC Scopus subject areas

  • Urology

Cite this

Regulation of androgen and vitamin D receptors by 1,25-dihydroxyvitamin D3 in human prostate epithelial and stromal cells. / Leman, Eddy S.; DeMiguel, Fernando; Gao, Allen C; Getzenberg, Robert H.

In: Journal of Urology, Vol. 170, No. 1, 01.07.2003, p. 235-240.

Research output: Contribution to journalArticle

Leman, Eddy S. ; DeMiguel, Fernando ; Gao, Allen C ; Getzenberg, Robert H. / Regulation of androgen and vitamin D receptors by 1,25-dihydroxyvitamin D3 in human prostate epithelial and stromal cells. In: Journal of Urology. 2003 ; Vol. 170, No. 1. pp. 235-240.
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N2 - Purpose: The mechanisms of the interaction between 1,25-dihydroxyvitamin D3 (1,25 D) and androgens, and their respective receptors in their action on the prostate are not completely understood. We examined the interplay of 1,25 D and androgens on the epithelial and stromal cells of the prostate. Materials and Methods: The human neonatal prostatic epithelial cell line 267B-1 (BRFF, Inc., Ijamsville, Maryland) and primary cultures of human prostate stromal cells were treated with medium containing 5 or 10 μM 1,25 D or ethanol (control) in the presence or absence of 10 nM dihydrotestosterone (DHT) (Sigma Chemical Co., St. Louis, Missouri). Protein levels of androgen receptor (AR) and vitamin D receptor (VDR) were determined by immunoblot analysis of whole cell extracts. Electrophoresis mobility shift assays were used to determine AR and VDR DNA binding activities. Results: The VDR protein level of 267B-1 cells was increased in the presence of 1,25 D (with the maximum effects seen at 24 hours) regardless of the presence or absence of DHT. In addition, exogenous DHT increased the AR and VDR DNA binding activities of 267B-1 and stromal cells in the presence of 1,25 D. Conclusions: ARs in the normal prostate are regulated by androgens, whereas VDRs in the normal prostate can be regulated by 1,25 D as well as by other androgens such as testosterone. This finding further supports the concept that 1,25 D as a steroid hormone, in addition to other androgens such as DHT, may have a role in the growth and differentiation of normal prostate.

AB - Purpose: The mechanisms of the interaction between 1,25-dihydroxyvitamin D3 (1,25 D) and androgens, and their respective receptors in their action on the prostate are not completely understood. We examined the interplay of 1,25 D and androgens on the epithelial and stromal cells of the prostate. Materials and Methods: The human neonatal prostatic epithelial cell line 267B-1 (BRFF, Inc., Ijamsville, Maryland) and primary cultures of human prostate stromal cells were treated with medium containing 5 or 10 μM 1,25 D or ethanol (control) in the presence or absence of 10 nM dihydrotestosterone (DHT) (Sigma Chemical Co., St. Louis, Missouri). Protein levels of androgen receptor (AR) and vitamin D receptor (VDR) were determined by immunoblot analysis of whole cell extracts. Electrophoresis mobility shift assays were used to determine AR and VDR DNA binding activities. Results: The VDR protein level of 267B-1 cells was increased in the presence of 1,25 D (with the maximum effects seen at 24 hours) regardless of the presence or absence of DHT. In addition, exogenous DHT increased the AR and VDR DNA binding activities of 267B-1 and stromal cells in the presence of 1,25 D. Conclusions: ARs in the normal prostate are regulated by androgens, whereas VDRs in the normal prostate can be regulated by 1,25 D as well as by other androgens such as testosterone. This finding further supports the concept that 1,25 D as a steroid hormone, in addition to other androgens such as DHT, may have a role in the growth and differentiation of normal prostate.

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