The Ah receptor inhibits estrogen-induced estrogen receptor β in breast cancer cells

doi:10.1016/j.bbrc.2004.05.132

Biochemical and Biophysical Research Communications

Volume 320, Issue 1, 16 July 2004, Pages 76-82

https://doi.org/10.1016/j.bbrc.2004.05.132 Get rights and content

Abstract

We have studied the effect of the aryl hydrocarbon receptor ligand 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on estrogen receptor (ER) β gene expression in the human breast cancer cell line, T47D. TCDD inhibited 17β-estradiol (E₂)-induced up-regulation of both ER β wild type and ER β cx mRNA. Cycloheximide pre-treatment had no inhibitory effect, and the estimated half-life of ER β mRNA of about 33 min was not changed by any hormone administration. Chromatin immunoprecipitation experiments showed recruitment of ER α to the ER β promoter. Gel mobility shift experiments revealed an E₂-induced protein binding to a half site estrogen response element in the ER β promoter, and TCDD reduced that binding. These results show that ER α regulates the expression of its own heterodimerization partner, ER β, in T47D cells. TCDD, an anti-estrogenic compound, inhibits ER α-mediated induction of ER β mRNA. These findings add to our understanding of cross talk between dioxin and estrogen signaling in human cells.

Section snippets

Materials and methods

Cell culture. The human breast cancer cells T47D (ATCC) were cultured in a 1:1 mixture of Ham's nutrient mixture F12 and DMEM (Gibco-BRL) supplemented with 10% fetal bovine serum (FBS) (Life Technologies) and 1% PEST (100 U penicillin/ml and 100 μg streptomycin/ml) (Gibco-BRL). The cells were seeded on plates 3 days before hormone treatment in a 1:1 mixture of phenol-red free DMEM (Gibco-BRL) and F12 containing 5% dextran-coated charcoal-treated FBS and 1% PEST (DCC-medium).

hRNA isolation, DNA

Inhibition of estrogen-induced ER β mRNA expression by TCDD

In order to determine the levels of ER β mRNA, we carried out QRT-PCR. By designing primers situated in exon 7 and 8, we were able to differentiate between ER β wt and the isoform ER β cx, since the structural difference between ER β wt and ER β cx is caused by an exchange in exon 8. The exon 8 is part of the ligand-binding domain of ER β. ER β cx does not bind ligand, but may inhibit ER α transactivation [9]. In our T47D cells, the constitutive level of ER β cx mRNA is about threefold higher

Acknowledgements

This research was supported by grants from the Swedish Cancer Fund and from KaroBio AB. S.K. holds an EC Marie Curie individual postdoctoral fellowship (MCFI-2000-02111).

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The Ah receptor inhibits estrogen-induced estrogen receptor β in breast cancer cells

Abstract

Section snippets

Materials and methods

Inhibition of estrogen-induced ER β mRNA expression by TCDD

Acknowledgements

FEBS Lett.

FEBS Lett.

Eur. J. Cancer

Cancer Lett.

J. Biol. Chem.

Arch. Biochem. Biophys.

Arch. Biochem. Biophys.

J. Biol. Chem.

J. Biol. Chem.

Cell

J. Biol. Chem.

J. Steroid Biochem. Mol. Biol.

J. Steroid Biochem. Mol. Biol.

Mol. Cell. Endocrinol.

FEBS Lett.

Mol. Cell. Endocrinol.

Prognostic value of steroid hormone receptors: multivariate analysis of systemically untreated patients with node negative primary breast cancer

Cancer Res.

Cloning of a novel receptor expressed in rat prostate and ovary

Proc. Natl. Acad. Sci. USA

Oestrogen receptor beta in breast cancer: good, bad or still too early to tell?

J. Pathol.

Selective loss of estrogen receptor beta in malignant human colon

Cancer Res.

Loss of estrogen receptor beta expression in malignant human prostate cells in primary cultures and in prostate cancer tissues

J. Clin. Endocrinol. Metab.

Molecular cloning and characterization of human estrogen receptor beta cx: a potential inhibitor of estrogen action in human

Nucleic Acids Res.

Increased expression of estrogen receptor beta mRNA in tamoxifen-resistant breast cancer patients

Cancer Res.