Down-regulation of the glucocorticoid receptor by dexamethasone in cultured human skin fibroblasts

Implications for the regulation of aromatase activity

Gary Berkovitz, K. M. Carter, C. J. Migeon, T. R. Brown

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Human genital skin fibroblasts grown in cell culture contain aromatase activity and glucocorticoid receptors, providing a model for studying the relationship between changes in levels of dexamethasone (DEX) receptor binding and changes in the response of aromatase to DEX. Incubation of cells with media containing DEX produced a time-dependent decline in receptor number, with a nadir at 18-24 h. When DEX was removed from the media, only 40-60% of the lost receptor content was recovered. Binding of DEX to its receptor declined in a dose-dependent fashion after 24-h exposure to this glucocorticoid; the concentration of DEX that produced a half-maximal decline (2.2 nmol/L) was in the same range as the K(d) for the receptor-DEX complex (14 nmol/L). Incubation of cells with DEX for 20 h also reduced the level of DEX receptor binding in purified nuclei and nuclear matrix by 76% and 89%, respectively. When subcellular fractions were prepared after incubation of cells with DEX for 1-24 h, whole cell, cytoslic and nuclear DEX receptor binding declined in parallel with time. Incubation of skin fibroblasts with DEX resulted in progressive stimulation of aromatase activity, with a peak at 24 h followed by a return to baseline at 72 h. The initial stimulation of aromatase was mediated by DEX receptors. However, the decline in aromatase activity after prolonged DEX exposure did not appear to be due to a decline in the level of DEX receptor binding. The data supporting this last conclusion included the following. When cells were washed free of DEX after 48 h, DEX receptor binding recovered within 24 h, whereas aromatase activity could not be maximally restimulated until 36 h; when cells were incubated with media containing DEX and cycloheximide for 1-48 h, DEX receptor binding declined to a nadir by 24 h, whereas aromatase activity rose continuously up to 48 h. These findings are consistent with the concept that the aromatase gene in skin fibroblasts is subject to both positive and negative regulation.

Original languageEnglish
Pages (from-to)1029-1036
Number of pages8
JournalJournal of Clinical Endocrinology and Metabolism
Volume66
Issue number5
StatePublished - Jan 1 1988
Externally publishedYes

Fingerprint

Aromatase
Glucocorticoid Receptors
Fibroblasts
Dexamethasone
Skin
Down-Regulation
Cells
Nuclear Matrix
Subcellular Fractions
dexamethasone receptor
Cycloheximide
Cytoplasmic and Nuclear Receptors
Cell culture
Glucocorticoids
Cell Culture Techniques
Genes

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology, Diabetes and Metabolism

Cite this

Down-regulation of the glucocorticoid receptor by dexamethasone in cultured human skin fibroblasts : Implications for the regulation of aromatase activity. / Berkovitz, Gary; Carter, K. M.; Migeon, C. J.; Brown, T. R.

In: Journal of Clinical Endocrinology and Metabolism, Vol. 66, No. 5, 01.01.1988, p. 1029-1036.

Research output: Contribution to journalArticle

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title = "Down-regulation of the glucocorticoid receptor by dexamethasone in cultured human skin fibroblasts: Implications for the regulation of aromatase activity",
abstract = "Human genital skin fibroblasts grown in cell culture contain aromatase activity and glucocorticoid receptors, providing a model for studying the relationship between changes in levels of dexamethasone (DEX) receptor binding and changes in the response of aromatase to DEX. Incubation of cells with media containing DEX produced a time-dependent decline in receptor number, with a nadir at 18-24 h. When DEX was removed from the media, only 40-60{\%} of the lost receptor content was recovered. Binding of DEX to its receptor declined in a dose-dependent fashion after 24-h exposure to this glucocorticoid; the concentration of DEX that produced a half-maximal decline (2.2 nmol/L) was in the same range as the K(d) for the receptor-DEX complex (14 nmol/L). Incubation of cells with DEX for 20 h also reduced the level of DEX receptor binding in purified nuclei and nuclear matrix by 76{\%} and 89{\%}, respectively. When subcellular fractions were prepared after incubation of cells with DEX for 1-24 h, whole cell, cytoslic and nuclear DEX receptor binding declined in parallel with time. Incubation of skin fibroblasts with DEX resulted in progressive stimulation of aromatase activity, with a peak at 24 h followed by a return to baseline at 72 h. The initial stimulation of aromatase was mediated by DEX receptors. However, the decline in aromatase activity after prolonged DEX exposure did not appear to be due to a decline in the level of DEX receptor binding. The data supporting this last conclusion included the following. When cells were washed free of DEX after 48 h, DEX receptor binding recovered within 24 h, whereas aromatase activity could not be maximally restimulated until 36 h; when cells were incubated with media containing DEX and cycloheximide for 1-48 h, DEX receptor binding declined to a nadir by 24 h, whereas aromatase activity rose continuously up to 48 h. These findings are consistent with the concept that the aromatase gene in skin fibroblasts is subject to both positive and negative regulation.",
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