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ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions

The two estrogen receptors ERα and ERβ are nuclear receptors that bind estrogen (E2) and function as ligand-inducible transcription factors. They are homologues and can form dimers with each other and bind to the same estrogen-response element motifs in the DNA. ERα drives breast cancer growth where...

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Autores principales: Song, Dandan, He, Huan, Indukuri, Rajitha, Huang, Zhiqiang, Stepanauskaite, Lina, Sinha, Indranil, Haldosén, Lars-Arne, Zhao, Chunyan, Williams, Cecilia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9299245/
https://www.ncbi.nlm.nih.gov/pubmed/35872983
http://dx.doi.org/10.3389/fendo.2022.930227
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author Song, Dandan
He, Huan
Indukuri, Rajitha
Huang, Zhiqiang
Stepanauskaite, Lina
Sinha, Indranil
Haldosén, Lars-Arne
Zhao, Chunyan
Williams, Cecilia
author_facet Song, Dandan
He, Huan
Indukuri, Rajitha
Huang, Zhiqiang
Stepanauskaite, Lina
Sinha, Indranil
Haldosén, Lars-Arne
Zhao, Chunyan
Williams, Cecilia
author_sort Song, Dandan
collection PubMed
description The two estrogen receptors ERα and ERβ are nuclear receptors that bind estrogen (E2) and function as ligand-inducible transcription factors. They are homologues and can form dimers with each other and bind to the same estrogen-response element motifs in the DNA. ERα drives breast cancer growth whereas ERβ has been reported to be anti-proliferative. However, they are rarely expressed in the same cells, and it is not fully investigated to which extent their functions are different because of inherent differences or because of different cellular context. To dissect their similarities and differences, we here generated a novel estrogen-dependent cell model where ERα homodimers can be directly compared to ERβ homodimers within the identical cellular context. By using CRISPR-cas9 to delete ERα in breast cancer MCF7 cells with Tet-Off-inducible ERβ expression, we generated MCF7 cells that express ERβ but not ERα. MCF7 (ERβ only) cells exhibited regulation of estrogen-responsive targets in a ligand-dependent manner. We demonstrated that either ER was required for MCF7 proliferation, but while E2 increased proliferation via ERα, it reduced proliferation through a G2/M arrest via ERβ. The two ERs also impacted migration differently. In absence of ligand, ERβ increased migration, but upon E2 treatment, ERβ reduced migration. E2 via ERα, on the other hand, had no significant impact on migration. RNA sequencing revealed that E2 regulated a transcriptome of around 800 genes via each receptor, but over half were specific for either ERα or ERβ (417 and 503 genes, respectively). Functional gene ontology enrichment analysis reinforced that E2 regulated cell proliferation in opposite directions depending on the ER, and that ERβ specifically impacted extracellular matrix organization. We corroborated that ERβ bound to cis-regulatory chromatin of its unique proposed migration-related direct targets ANXA9 and TFAP2C. In conclusion, we demonstrate that within the same cellular context, the two ERs regulate cell proliferation in the opposite manner, impact migration differently, and each receptor also regulates a distinct set of target genes in response to E2. The developed cell model provides a novel and valuable resource to further complement the mechanistic understanding of the two different ER isoforms.
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spelling pubmed-92992452022-07-21 ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions Song, Dandan He, Huan Indukuri, Rajitha Huang, Zhiqiang Stepanauskaite, Lina Sinha, Indranil Haldosén, Lars-Arne Zhao, Chunyan Williams, Cecilia Front Endocrinol (Lausanne) Endocrinology The two estrogen receptors ERα and ERβ are nuclear receptors that bind estrogen (E2) and function as ligand-inducible transcription factors. They are homologues and can form dimers with each other and bind to the same estrogen-response element motifs in the DNA. ERα drives breast cancer growth whereas ERβ has been reported to be anti-proliferative. However, they are rarely expressed in the same cells, and it is not fully investigated to which extent their functions are different because of inherent differences or because of different cellular context. To dissect their similarities and differences, we here generated a novel estrogen-dependent cell model where ERα homodimers can be directly compared to ERβ homodimers within the identical cellular context. By using CRISPR-cas9 to delete ERα in breast cancer MCF7 cells with Tet-Off-inducible ERβ expression, we generated MCF7 cells that express ERβ but not ERα. MCF7 (ERβ only) cells exhibited regulation of estrogen-responsive targets in a ligand-dependent manner. We demonstrated that either ER was required for MCF7 proliferation, but while E2 increased proliferation via ERα, it reduced proliferation through a G2/M arrest via ERβ. The two ERs also impacted migration differently. In absence of ligand, ERβ increased migration, but upon E2 treatment, ERβ reduced migration. E2 via ERα, on the other hand, had no significant impact on migration. RNA sequencing revealed that E2 regulated a transcriptome of around 800 genes via each receptor, but over half were specific for either ERα or ERβ (417 and 503 genes, respectively). Functional gene ontology enrichment analysis reinforced that E2 regulated cell proliferation in opposite directions depending on the ER, and that ERβ specifically impacted extracellular matrix organization. We corroborated that ERβ bound to cis-regulatory chromatin of its unique proposed migration-related direct targets ANXA9 and TFAP2C. In conclusion, we demonstrate that within the same cellular context, the two ERs regulate cell proliferation in the opposite manner, impact migration differently, and each receptor also regulates a distinct set of target genes in response to E2. The developed cell model provides a novel and valuable resource to further complement the mechanistic understanding of the two different ER isoforms. Frontiers Media S.A. 2022-07-06 /pmc/articles/PMC9299245/ /pubmed/35872983 http://dx.doi.org/10.3389/fendo.2022.930227 Text en Copyright © 2022 Song, He, Indukuri, Huang, Stepanauskaite, Sinha, Haldosén, Zhao and Williams https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Endocrinology
Song, Dandan
He, Huan
Indukuri, Rajitha
Huang, Zhiqiang
Stepanauskaite, Lina
Sinha, Indranil
Haldosén, Lars-Arne
Zhao, Chunyan
Williams, Cecilia
ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions
title ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions
title_full ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions
title_fullStr ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions
title_full_unstemmed ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions
title_short ERα and ERβ Homodimers in the Same Cellular Context Regulate Distinct Transcriptomes and Functions
title_sort erα and erβ homodimers in the same cellular context regulate distinct transcriptomes and functions
topic Endocrinology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9299245/
https://www.ncbi.nlm.nih.gov/pubmed/35872983
http://dx.doi.org/10.3389/fendo.2022.930227
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