Regulation of estrogen receptor-α gene expression by 1,25- dihydroxyvitamin D in MCF-7 cells

Adriana Stoica, Miguel Saceda, Amina Fakhro, Harrison B. Solomon, Bradley D. Fenster, Mary Beth Martin

Research output: Contribution to journalArticle

65 Citations (Scopus)

Abstract

This report describes an investigation of the role of 1,25- dihydroxyvitamin D (VD3) in the regulation of estrogen receptor-α (ER) in the ER-positive breast cancer cell line, MCF-7. Treatment of cells with 10 nM VD3 resulted in a 50% decline in the concentration of ER protein at 24 h. Scatchard analysis showed a corresponding decrease in the number of estradiol binding sites and no alteration in the binding affinity of estradiol for the ER (K(d) = 0.08 nM in VD3-treated cells compared with K(d) = 0.07 nM in control cells). Vitamin D treatment also caused a 50% decrease in the steady state amount of ER mRNA, which was maximal by 18 h. In vitro transcription run-on experiments demonstrated a decrease of approximately 60% in transcription of the estrogen receptor gene. Transient transfections using an ER promoter-CAT construct also demonstrated a 40% decrease in CAT activity after VD3 treatment. Sequence analysis identified a potential vitamin D response element (nVDRE) within the ER promoter. When this element was mutated, the ability of VD3 to block transcription from the ER promoter was lost. When the nVDRE was placed upstream of a heterologous promoter, nVDRE- SV40-CAT, treatment with VD3 resulted in a 50% decrease in CAT activity. Interestingly, co-transfection of either the ER promoter-CAT or the nVDRE- SV40-CAT construct and a vitamin D receptor expression vector into COS-1 or CV-1 cells showed an approximately 4-fold increase in CAT activity after VD3 treatment. Taken together these data suggest that VD3 inhibition of ER gene transcription is mediated through a nVDRE in the ER promoter. Inhibition appears to be cell specific.

Original languageEnglish
Pages (from-to)640-651
Number of pages12
JournalJournal of Cellular Biochemistry
Volume75
Issue number4
DOIs
Publication statusPublished - 15 Dec 1999
Externally publishedYes

Fingerprint

MCF-7 Cells
Gene expression
Estrogen Receptors
Gene Expression
Transcription
Cells
Transfection
Estradiol
Vitamin D Response Element
1,25-dihydroxyvitamin D
Genes
Estradiol Receptors
Calcitriol Receptors
Vitamin D
Sequence Analysis
Binding Sites
Breast Neoplasms
Cell Line
Messenger RNA

Keywords

  • 1,25-dihydroxyvitamin 3
  • ER
  • MCF-7 cells

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology

Cite this

Regulation of estrogen receptor-α gene expression by 1,25- dihydroxyvitamin D in MCF-7 cells. / Stoica, Adriana; Saceda, Miguel; Fakhro, Amina; Solomon, Harrison B.; Fenster, Bradley D.; Martin, Mary Beth.

In: Journal of Cellular Biochemistry, Vol. 75, No. 4, 15.12.1999, p. 640-651.

Research output: Contribution to journalArticle

Stoica, Adriana ; Saceda, Miguel ; Fakhro, Amina ; Solomon, Harrison B. ; Fenster, Bradley D. ; Martin, Mary Beth. / Regulation of estrogen receptor-α gene expression by 1,25- dihydroxyvitamin D in MCF-7 cells. In: Journal of Cellular Biochemistry. 1999 ; Vol. 75, No. 4. pp. 640-651.
@article{7cd87098f9cd4adea772f367ac0cc439,
title = "Regulation of estrogen receptor-α gene expression by 1,25- dihydroxyvitamin D in MCF-7 cells",
abstract = "This report describes an investigation of the role of 1,25- dihydroxyvitamin D (VD3) in the regulation of estrogen receptor-α (ER) in the ER-positive breast cancer cell line, MCF-7. Treatment of cells with 10 nM VD3 resulted in a 50{\%} decline in the concentration of ER protein at 24 h. Scatchard analysis showed a corresponding decrease in the number of estradiol binding sites and no alteration in the binding affinity of estradiol for the ER (K(d) = 0.08 nM in VD3-treated cells compared with K(d) = 0.07 nM in control cells). Vitamin D treatment also caused a 50{\%} decrease in the steady state amount of ER mRNA, which was maximal by 18 h. In vitro transcription run-on experiments demonstrated a decrease of approximately 60{\%} in transcription of the estrogen receptor gene. Transient transfections using an ER promoter-CAT construct also demonstrated a 40{\%} decrease in CAT activity after VD3 treatment. Sequence analysis identified a potential vitamin D response element (nVDRE) within the ER promoter. When this element was mutated, the ability of VD3 to block transcription from the ER promoter was lost. When the nVDRE was placed upstream of a heterologous promoter, nVDRE- SV40-CAT, treatment with VD3 resulted in a 50{\%} decrease in CAT activity. Interestingly, co-transfection of either the ER promoter-CAT or the nVDRE- SV40-CAT construct and a vitamin D receptor expression vector into COS-1 or CV-1 cells showed an approximately 4-fold increase in CAT activity after VD3 treatment. Taken together these data suggest that VD3 inhibition of ER gene transcription is mediated through a nVDRE in the ER promoter. Inhibition appears to be cell specific.",
keywords = "1,25-dihydroxyvitamin 3, ER, MCF-7 cells",
author = "Adriana Stoica and Miguel Saceda and Amina Fakhro and Solomon, {Harrison B.} and Fenster, {Bradley D.} and Martin, {Mary Beth}",
year = "1999",
month = "12",
day = "15",
doi = "10.1002/(SICI)1097-4644(19991215)75:4<640::AID-JCB10>3.0.CO;2-8",
language = "English",
volume = "75",
pages = "640--651",
journal = "Journal of Cellular Biochemistry",
issn = "0730-2312",
publisher = "Wiley-Liss Inc.",
number = "4",

}

TY - JOUR

T1 - Regulation of estrogen receptor-α gene expression by 1,25- dihydroxyvitamin D in MCF-7 cells

AU - Stoica, Adriana

AU - Saceda, Miguel

AU - Fakhro, Amina

AU - Solomon, Harrison B.

AU - Fenster, Bradley D.

AU - Martin, Mary Beth

PY - 1999/12/15

Y1 - 1999/12/15

N2 - This report describes an investigation of the role of 1,25- dihydroxyvitamin D (VD3) in the regulation of estrogen receptor-α (ER) in the ER-positive breast cancer cell line, MCF-7. Treatment of cells with 10 nM VD3 resulted in a 50% decline in the concentration of ER protein at 24 h. Scatchard analysis showed a corresponding decrease in the number of estradiol binding sites and no alteration in the binding affinity of estradiol for the ER (K(d) = 0.08 nM in VD3-treated cells compared with K(d) = 0.07 nM in control cells). Vitamin D treatment also caused a 50% decrease in the steady state amount of ER mRNA, which was maximal by 18 h. In vitro transcription run-on experiments demonstrated a decrease of approximately 60% in transcription of the estrogen receptor gene. Transient transfections using an ER promoter-CAT construct also demonstrated a 40% decrease in CAT activity after VD3 treatment. Sequence analysis identified a potential vitamin D response element (nVDRE) within the ER promoter. When this element was mutated, the ability of VD3 to block transcription from the ER promoter was lost. When the nVDRE was placed upstream of a heterologous promoter, nVDRE- SV40-CAT, treatment with VD3 resulted in a 50% decrease in CAT activity. Interestingly, co-transfection of either the ER promoter-CAT or the nVDRE- SV40-CAT construct and a vitamin D receptor expression vector into COS-1 or CV-1 cells showed an approximately 4-fold increase in CAT activity after VD3 treatment. Taken together these data suggest that VD3 inhibition of ER gene transcription is mediated through a nVDRE in the ER promoter. Inhibition appears to be cell specific.

AB - This report describes an investigation of the role of 1,25- dihydroxyvitamin D (VD3) in the regulation of estrogen receptor-α (ER) in the ER-positive breast cancer cell line, MCF-7. Treatment of cells with 10 nM VD3 resulted in a 50% decline in the concentration of ER protein at 24 h. Scatchard analysis showed a corresponding decrease in the number of estradiol binding sites and no alteration in the binding affinity of estradiol for the ER (K(d) = 0.08 nM in VD3-treated cells compared with K(d) = 0.07 nM in control cells). Vitamin D treatment also caused a 50% decrease in the steady state amount of ER mRNA, which was maximal by 18 h. In vitro transcription run-on experiments demonstrated a decrease of approximately 60% in transcription of the estrogen receptor gene. Transient transfections using an ER promoter-CAT construct also demonstrated a 40% decrease in CAT activity after VD3 treatment. Sequence analysis identified a potential vitamin D response element (nVDRE) within the ER promoter. When this element was mutated, the ability of VD3 to block transcription from the ER promoter was lost. When the nVDRE was placed upstream of a heterologous promoter, nVDRE- SV40-CAT, treatment with VD3 resulted in a 50% decrease in CAT activity. Interestingly, co-transfection of either the ER promoter-CAT or the nVDRE- SV40-CAT construct and a vitamin D receptor expression vector into COS-1 or CV-1 cells showed an approximately 4-fold increase in CAT activity after VD3 treatment. Taken together these data suggest that VD3 inhibition of ER gene transcription is mediated through a nVDRE in the ER promoter. Inhibition appears to be cell specific.

KW - 1,25-dihydroxyvitamin 3

KW - ER

KW - MCF-7 cells

UR - http://www.scopus.com/inward/record.url?scp=0033573305&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0033573305&partnerID=8YFLogxK

U2 - 10.1002/(SICI)1097-4644(19991215)75:4<640::AID-JCB10>3.0.CO;2-8

DO - 10.1002/(SICI)1097-4644(19991215)75:4<640::AID-JCB10>3.0.CO;2-8

M3 - Article

C2 - 10572247

AN - SCOPUS:0033573305

VL - 75

SP - 640

EP - 651

JO - Journal of Cellular Biochemistry

JF - Journal of Cellular Biochemistry

SN - 0730-2312

IS - 4

ER -