Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP

Y. Gong, L. J. Blok, J. E. Perry, J. K. Lindzey, D. J. Tindall

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

Elevation of intracellular calcium levels in the presence of normal androgen levels has been implicated in apoptotic prostate cell death. Since the androgen receptor (AR) plays a critical role in the regulation of growth and differentiation of the prostate, it was of interest to determine whether Ca2+ would affect the expression of androgen receptor messenger RNA (mRNA) and protein, thus affecting the ability of androgens to control prostate function. AR-positive human prostate cancer cells, LNCaP, were incubated with either the calcium ionophore A23187 or the intracellular endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin. Subsequently, AR mRNA and protein levels were assessed by Northern and Western blot analysis. Both A23187 and thapsigargin were found to down-regulate steady state AR mRNA levels in a time- and dose-dependent manner. AR mRNA began to decrease after 6-8 h of incubation with 10-6 M A23187 or 10-7 M thapsigargin, reaching a nadir at 16 and 10 h of incubation, respectively. In contrast, control mRNA (glyceraldehyde 3-phosphate dehydrogenase) did not change significantly during the treatments with either A23187 or thapsigargin. AR protein levels were found to be decreased after 12 h of incubation with either 10-6 M A23187 or 10-7 M thapsigargin. The decrease in AR mRNA and protein seemed to precede apoptosis, since neither A23187 (24 h) nor thapsigargin (30 h) was found to alter cell morphology within the treatment time. Cycloheximide and actinomycin D were unable to change the calcium-mediated decrease in AR mRNA, ruling out the necessity for de novo protein synthesis or a change in mRNA stability. Moreover, the decrease in AR mRNA induced by calcium does not seem to involve protein kinase C- or calmodulin-dependent pathways, since inhibitors of these cellular components had no effect. Nuclear run-on assays demonstrated little or no effects of either A23187 or thapsigargin treatment on AR gene transcription (8 h and 10 h). In conclusion, these studies show that intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.

Original languageEnglish (US)
Pages (from-to)2172-2178
Number of pages7
JournalEndocrinology
Volume136
Issue number5
StatePublished - 1995

Fingerprint

Androgen Receptors
Prostatic Neoplasms
Thapsigargin
Calcimycin
Calcium
Cell Line
Messenger RNA
Prostate
Proteins
Androgens
Glyceraldehyde-3-Phosphate Dehydrogenases
Calcium-Transporting ATPases
Calcium Ionophores
RNA Stability
Dactinomycin
Calmodulin
Cycloheximide
Endoplasmic Reticulum
Northern Blotting
Protein Kinase C

ASJC Scopus subject areas

  • Endocrinology
  • Endocrinology, Diabetes and Metabolism

Cite this

Gong, Y., Blok, L. J., Perry, J. E., Lindzey, J. K., & Tindall, D. J. (1995). Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP. Endocrinology, 136(5), 2172-2178.

Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP. / Gong, Y.; Blok, L. J.; Perry, J. E.; Lindzey, J. K.; Tindall, D. J.

In: Endocrinology, Vol. 136, No. 5, 1995, p. 2172-2178.

Research output: Contribution to journalArticle

Gong, Y, Blok, LJ, Perry, JE, Lindzey, JK & Tindall, DJ 1995, 'Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP', Endocrinology, vol. 136, no. 5, pp. 2172-2178.
Gong Y, Blok LJ, Perry JE, Lindzey JK, Tindall DJ. Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP. Endocrinology. 1995;136(5):2172-2178.
Gong, Y. ; Blok, L. J. ; Perry, J. E. ; Lindzey, J. K. ; Tindall, D. J. / Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP. In: Endocrinology. 1995 ; Vol. 136, No. 5. pp. 2172-2178.
@article{faae3979acc84bbe8fb4aceb46bf3b6b,
title = "Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP",
abstract = "Elevation of intracellular calcium levels in the presence of normal androgen levels has been implicated in apoptotic prostate cell death. Since the androgen receptor (AR) plays a critical role in the regulation of growth and differentiation of the prostate, it was of interest to determine whether Ca2+ would affect the expression of androgen receptor messenger RNA (mRNA) and protein, thus affecting the ability of androgens to control prostate function. AR-positive human prostate cancer cells, LNCaP, were incubated with either the calcium ionophore A23187 or the intracellular endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin. Subsequently, AR mRNA and protein levels were assessed by Northern and Western blot analysis. Both A23187 and thapsigargin were found to down-regulate steady state AR mRNA levels in a time- and dose-dependent manner. AR mRNA began to decrease after 6-8 h of incubation with 10-6 M A23187 or 10-7 M thapsigargin, reaching a nadir at 16 and 10 h of incubation, respectively. In contrast, control mRNA (glyceraldehyde 3-phosphate dehydrogenase) did not change significantly during the treatments with either A23187 or thapsigargin. AR protein levels were found to be decreased after 12 h of incubation with either 10-6 M A23187 or 10-7 M thapsigargin. The decrease in AR mRNA and protein seemed to precede apoptosis, since neither A23187 (24 h) nor thapsigargin (30 h) was found to alter cell morphology within the treatment time. Cycloheximide and actinomycin D were unable to change the calcium-mediated decrease in AR mRNA, ruling out the necessity for de novo protein synthesis or a change in mRNA stability. Moreover, the decrease in AR mRNA induced by calcium does not seem to involve protein kinase C- or calmodulin-dependent pathways, since inhibitors of these cellular components had no effect. Nuclear run-on assays demonstrated little or no effects of either A23187 or thapsigargin treatment on AR gene transcription (8 h and 10 h). In conclusion, these studies show that intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.",
author = "Y. Gong and Blok, {L. J.} and Perry, {J. E.} and Lindzey, {J. K.} and Tindall, {D. J.}",
year = "1995",
language = "English (US)",
volume = "136",
pages = "2172--2178",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "5",

}

TY - JOUR

T1 - Calcium regulation of androgen receptor expression in the human prostate cancer cell line LNCaP

AU - Gong, Y.

AU - Blok, L. J.

AU - Perry, J. E.

AU - Lindzey, J. K.

AU - Tindall, D. J.

PY - 1995

Y1 - 1995

N2 - Elevation of intracellular calcium levels in the presence of normal androgen levels has been implicated in apoptotic prostate cell death. Since the androgen receptor (AR) plays a critical role in the regulation of growth and differentiation of the prostate, it was of interest to determine whether Ca2+ would affect the expression of androgen receptor messenger RNA (mRNA) and protein, thus affecting the ability of androgens to control prostate function. AR-positive human prostate cancer cells, LNCaP, were incubated with either the calcium ionophore A23187 or the intracellular endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin. Subsequently, AR mRNA and protein levels were assessed by Northern and Western blot analysis. Both A23187 and thapsigargin were found to down-regulate steady state AR mRNA levels in a time- and dose-dependent manner. AR mRNA began to decrease after 6-8 h of incubation with 10-6 M A23187 or 10-7 M thapsigargin, reaching a nadir at 16 and 10 h of incubation, respectively. In contrast, control mRNA (glyceraldehyde 3-phosphate dehydrogenase) did not change significantly during the treatments with either A23187 or thapsigargin. AR protein levels were found to be decreased after 12 h of incubation with either 10-6 M A23187 or 10-7 M thapsigargin. The decrease in AR mRNA and protein seemed to precede apoptosis, since neither A23187 (24 h) nor thapsigargin (30 h) was found to alter cell morphology within the treatment time. Cycloheximide and actinomycin D were unable to change the calcium-mediated decrease in AR mRNA, ruling out the necessity for de novo protein synthesis or a change in mRNA stability. Moreover, the decrease in AR mRNA induced by calcium does not seem to involve protein kinase C- or calmodulin-dependent pathways, since inhibitors of these cellular components had no effect. Nuclear run-on assays demonstrated little or no effects of either A23187 or thapsigargin treatment on AR gene transcription (8 h and 10 h). In conclusion, these studies show that intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.

AB - Elevation of intracellular calcium levels in the presence of normal androgen levels has been implicated in apoptotic prostate cell death. Since the androgen receptor (AR) plays a critical role in the regulation of growth and differentiation of the prostate, it was of interest to determine whether Ca2+ would affect the expression of androgen receptor messenger RNA (mRNA) and protein, thus affecting the ability of androgens to control prostate function. AR-positive human prostate cancer cells, LNCaP, were incubated with either the calcium ionophore A23187 or the intracellular endoplasmic reticulum Ca2+-ATPase inhibitor thapsigargin. Subsequently, AR mRNA and protein levels were assessed by Northern and Western blot analysis. Both A23187 and thapsigargin were found to down-regulate steady state AR mRNA levels in a time- and dose-dependent manner. AR mRNA began to decrease after 6-8 h of incubation with 10-6 M A23187 or 10-7 M thapsigargin, reaching a nadir at 16 and 10 h of incubation, respectively. In contrast, control mRNA (glyceraldehyde 3-phosphate dehydrogenase) did not change significantly during the treatments with either A23187 or thapsigargin. AR protein levels were found to be decreased after 12 h of incubation with either 10-6 M A23187 or 10-7 M thapsigargin. The decrease in AR mRNA and protein seemed to precede apoptosis, since neither A23187 (24 h) nor thapsigargin (30 h) was found to alter cell morphology within the treatment time. Cycloheximide and actinomycin D were unable to change the calcium-mediated decrease in AR mRNA, ruling out the necessity for de novo protein synthesis or a change in mRNA stability. Moreover, the decrease in AR mRNA induced by calcium does not seem to involve protein kinase C- or calmodulin-dependent pathways, since inhibitors of these cellular components had no effect. Nuclear run-on assays demonstrated little or no effects of either A23187 or thapsigargin treatment on AR gene transcription (8 h and 10 h). In conclusion, these studies show that intracellular calcium seems to be a potent regulator of AR gene expression in LNCaP cells.

UR - http://www.scopus.com/inward/record.url?scp=0028919447&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0028919447&partnerID=8YFLogxK

M3 - Article

C2 - 7720667

AN - SCOPUS:0028919447

VL - 136

SP - 2172

EP - 2178

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 5

ER -