Regulation of progesterone receptor messenger ribonucleic acid and protein levels in MCF-7 cells by estradiol: Analysis of estrogen’s effect on progesterone receptor synthesis and degradation

Ann M. Nardulli, Geoffrey L. Greene, Bert W. O’Malley, Benita S. Katzenellenbogen

Research output: Contribution to journalArticle

Abstract

The human breast cancer cell line MCF-7 responds to estrogens with increased progesterone receptor (PR) levels. In this study, we use dense amino acid density shift analyses to address directly the question of whether estrogen increases PR levels in MCF-7 cells by altering rates of receptor synthesis and/or degradation. Using different concentrations of estradiol (E2), which achieve PR levels that are half-maximal (3 × 10-11 M E2) or maximal (6 × 10-11 M E2), we have done sucrose gradient density shift analyses using dense (15N, 13C, 2H) amino acid incorporation to study rates of PR synthesis and degradation. These studies reveal a nonlinear loss of preexisting normal density receptor with time. From kinetic modeling analyses, equivalent rates of degradation are estimated for PR whether maximal or half-maximal levels are maintained, indicating that the major effect of E2 on PR content is to increase the rate of PR synthesis while leaving the degradation rate unaltered. The E2-stimulated increase in PR protein is also associated with increased levels of PR mRNA, as demonstrated by the use of a human PR cDNA probe. These density shift data provide evidence that the increased PR levels after estrogen exposure in MCF-7 cells are the result of an increased rate of receptor synthesis, rather than modulation of the rate of receptor degradation.

Original languageEnglish (US)
Pages (from-to)935-944
Number of pages10
JournalEndocrinology
Volume122
Issue number3
DOIs
StatePublished - Mar 1 1988

Fingerprint

MCF-7 Cells
Progesterone Receptors
Estradiol
Estrogens
RNA
Proteins
Amino Acids
Sucrose
Complementary DNA
Breast Neoplasms
Cell Line
Messenger RNA

ASJC Scopus subject areas

  • Endocrinology

Cite this

Regulation of progesterone receptor messenger ribonucleic acid and protein levels in MCF-7 cells by estradiol : Analysis of estrogen’s effect on progesterone receptor synthesis and degradation. / Nardulli, Ann M.; Greene, Geoffrey L.; O’Malley, Bert W.; Katzenellenbogen, Benita S.

In: Endocrinology, Vol. 122, No. 3, 01.03.1988, p. 935-944.

Research output: Contribution to journalArticle

@article{6f4233252b3243338a1b28fe6ee2d65d,
title = "Regulation of progesterone receptor messenger ribonucleic acid and protein levels in MCF-7 cells by estradiol: Analysis of estrogen’s effect on progesterone receptor synthesis and degradation",
abstract = "The human breast cancer cell line MCF-7 responds to estrogens with increased progesterone receptor (PR) levels. In this study, we use dense amino acid density shift analyses to address directly the question of whether estrogen increases PR levels in MCF-7 cells by altering rates of receptor synthesis and/or degradation. Using different concentrations of estradiol (E2), which achieve PR levels that are half-maximal (3 × 10-11 M E2) or maximal (6 × 10-11 M E2), we have done sucrose gradient density shift analyses using dense (15N, 13C, 2H) amino acid incorporation to study rates of PR synthesis and degradation. These studies reveal a nonlinear loss of preexisting normal density receptor with time. From kinetic modeling analyses, equivalent rates of degradation are estimated for PR whether maximal or half-maximal levels are maintained, indicating that the major effect of E2 on PR content is to increase the rate of PR synthesis while leaving the degradation rate unaltered. The E2-stimulated increase in PR protein is also associated with increased levels of PR mRNA, as demonstrated by the use of a human PR cDNA probe. These density shift data provide evidence that the increased PR levels after estrogen exposure in MCF-7 cells are the result of an increased rate of receptor synthesis, rather than modulation of the rate of receptor degradation.",
author = "Nardulli, {Ann M.} and Greene, {Geoffrey L.} and O’Malley, {Bert W.} and Katzenellenbogen, {Benita S.}",
year = "1988",
month = "3",
day = "1",
doi = "10.1210/endo-122-3-935",
language = "English (US)",
volume = "122",
pages = "935--944",
journal = "Endocrinology",
issn = "0013-7227",
publisher = "The Endocrine Society",
number = "3",

}

TY - JOUR

T1 - Regulation of progesterone receptor messenger ribonucleic acid and protein levels in MCF-7 cells by estradiol

T2 - Analysis of estrogen’s effect on progesterone receptor synthesis and degradation

AU - Nardulli, Ann M.

AU - Greene, Geoffrey L.

AU - O’Malley, Bert W.

AU - Katzenellenbogen, Benita S.

PY - 1988/3/1

Y1 - 1988/3/1

N2 - The human breast cancer cell line MCF-7 responds to estrogens with increased progesterone receptor (PR) levels. In this study, we use dense amino acid density shift analyses to address directly the question of whether estrogen increases PR levels in MCF-7 cells by altering rates of receptor synthesis and/or degradation. Using different concentrations of estradiol (E2), which achieve PR levels that are half-maximal (3 × 10-11 M E2) or maximal (6 × 10-11 M E2), we have done sucrose gradient density shift analyses using dense (15N, 13C, 2H) amino acid incorporation to study rates of PR synthesis and degradation. These studies reveal a nonlinear loss of preexisting normal density receptor with time. From kinetic modeling analyses, equivalent rates of degradation are estimated for PR whether maximal or half-maximal levels are maintained, indicating that the major effect of E2 on PR content is to increase the rate of PR synthesis while leaving the degradation rate unaltered. The E2-stimulated increase in PR protein is also associated with increased levels of PR mRNA, as demonstrated by the use of a human PR cDNA probe. These density shift data provide evidence that the increased PR levels after estrogen exposure in MCF-7 cells are the result of an increased rate of receptor synthesis, rather than modulation of the rate of receptor degradation.

AB - The human breast cancer cell line MCF-7 responds to estrogens with increased progesterone receptor (PR) levels. In this study, we use dense amino acid density shift analyses to address directly the question of whether estrogen increases PR levels in MCF-7 cells by altering rates of receptor synthesis and/or degradation. Using different concentrations of estradiol (E2), which achieve PR levels that are half-maximal (3 × 10-11 M E2) or maximal (6 × 10-11 M E2), we have done sucrose gradient density shift analyses using dense (15N, 13C, 2H) amino acid incorporation to study rates of PR synthesis and degradation. These studies reveal a nonlinear loss of preexisting normal density receptor with time. From kinetic modeling analyses, equivalent rates of degradation are estimated for PR whether maximal or half-maximal levels are maintained, indicating that the major effect of E2 on PR content is to increase the rate of PR synthesis while leaving the degradation rate unaltered. The E2-stimulated increase in PR protein is also associated with increased levels of PR mRNA, as demonstrated by the use of a human PR cDNA probe. These density shift data provide evidence that the increased PR levels after estrogen exposure in MCF-7 cells are the result of an increased rate of receptor synthesis, rather than modulation of the rate of receptor degradation.

UR - http://www.scopus.com/inward/record.url?scp=0023926059&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023926059&partnerID=8YFLogxK

U2 - 10.1210/endo-122-3-935

DO - 10.1210/endo-122-3-935

M3 - Article

C2 - 3342760

AN - SCOPUS:0023926059

VL - 122

SP - 935

EP - 944

JO - Endocrinology

JF - Endocrinology

SN - 0013-7227

IS - 3

ER -