Estrogen receptor α (ERα) mediates stimulatory effects of estrogen on aromatase (CYP19) gene expression in human placenta

Premlata Kumar, Amrita Kamat, Carole R. Mendelson

Research output: Contribution to journalArticle

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Abstract

A 246-bp region upstream of placenta-specific exon I.1 of the human aromatase (hCYP19) gene mediates placenta-specific, developmental, and O 2 regulation of expression. In this study, trophoblast differentiation and associated induction of CYP19 expression were prevented when cytotrophoblasts were cultured in phenol red-free medium containing charcoal-stripped serum or with the estrogen receptor (ER) antagonist, ICI 182,780, suggesting a stimulatory role of estrogen/ER. ERα protein was expressed in human trophoblasts and increased during syncytiotrophoblast differentiation, whereas ERα was undetectable. Mutational analysis revealed that an estrogen response element-like sequence (ERE-LS) at -208 bp is required for inductive effects of estradiol/ERα on hCYP19I.1 promoter activity in transfected COS-7 cells. Increased binding of syncytiotrophoblast compared with cytotrophoblast nuclear proteins to the ERE-LS was observed in vitro; however, ERα antibodies failed to supershift the complex and in vitro-transcribed/translated ERα did not bind. Nonetheless, chromatin immunoprecipitation assays in cultured trophoblasts revealed recruitment of endogenous ERα to the -255- to -155-bp region containing the ERE-LS before induction of hCYP19 expression; this was inhibited by ICI 182,780. Chromatin immunoprecipitation also revealed increased acetylated histone H3(K9/14) and decreased methylated histone H3(K9) associated with this region during trophoblast differentiation. These modifications were prevented when trophoblasts were incubated with ICI 182,780, suggesting that ERαrecruitment to the -255- to -155-bp region promotes histone modifications leading to increased hCYP19 transcription. Thus, during trophoblast differentiation, estrogen/ERα exerts a positive feedback role, which promotes permissive histone modifications that are associated with induction of hCYP19 gene transcription.

Original languageEnglish (US)
Pages (from-to)784-793
Number of pages10
JournalMolecular Endocrinology
Volume23
Issue number6
DOIs
StatePublished - Jun 2009

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Aromatase
Trophoblasts
Estrogen Receptors
Placenta
Estrogens
Gene Expression
Response Elements
Histone Code
Chromatin Immunoprecipitation
Histones
Phenolsulfonphthalein
Estradiol Receptors
Charcoal
COS Cells
Nuclear Proteins
Genes
Exons

ASJC Scopus subject areas

  • Molecular Biology
  • Endocrinology

Cite this

Estrogen receptor α (ERα) mediates stimulatory effects of estrogen on aromatase (CYP19) gene expression in human placenta. / Kumar, Premlata; Kamat, Amrita; Mendelson, Carole R.

In: Molecular Endocrinology, Vol. 23, No. 6, 06.2009, p. 784-793.

Research output: Contribution to journalArticle

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abstract = "A 246-bp region upstream of placenta-specific exon I.1 of the human aromatase (hCYP19) gene mediates placenta-specific, developmental, and O 2 regulation of expression. In this study, trophoblast differentiation and associated induction of CYP19 expression were prevented when cytotrophoblasts were cultured in phenol red-free medium containing charcoal-stripped serum or with the estrogen receptor (ER) antagonist, ICI 182,780, suggesting a stimulatory role of estrogen/ER. ERα protein was expressed in human trophoblasts and increased during syncytiotrophoblast differentiation, whereas ERα was undetectable. Mutational analysis revealed that an estrogen response element-like sequence (ERE-LS) at -208 bp is required for inductive effects of estradiol/ERα on hCYP19I.1 promoter activity in transfected COS-7 cells. Increased binding of syncytiotrophoblast compared with cytotrophoblast nuclear proteins to the ERE-LS was observed in vitro; however, ERα antibodies failed to supershift the complex and in vitro-transcribed/translated ERα did not bind. Nonetheless, chromatin immunoprecipitation assays in cultured trophoblasts revealed recruitment of endogenous ERα to the -255- to -155-bp region containing the ERE-LS before induction of hCYP19 expression; this was inhibited by ICI 182,780. Chromatin immunoprecipitation also revealed increased acetylated histone H3(K9/14) and decreased methylated histone H3(K9) associated with this region during trophoblast differentiation. These modifications were prevented when trophoblasts were incubated with ICI 182,780, suggesting that ERαrecruitment to the -255- to -155-bp region promotes histone modifications leading to increased hCYP19 transcription. Thus, during trophoblast differentiation, estrogen/ERα exerts a positive feedback role, which promotes permissive histone modifications that are associated with induction of hCYP19 gene transcription.",
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