Cited by 12Endometriosis Research and TreatmentTGF-β signaling in diseasesEstrogen and related hormone effects

Smad3/4 Binding to Promoter II of P450arom So As to Regulate Aromatase Expression in Endometriosis

Qu, Juan, Zhu, Yuanyuan, Wu, Xiadi, Zheng, Juan, Hou, Zhen, Cui, Yugui, Mao, Yundong, Liu, Jiayin·2017·Reproductive Sciences

Abstract

Activin A can stimulate aromatase P450 (P450arom) expression in eutopic endometrial stromal cells (ESCs) of endometriosis by activin type I receptor-Smad pathway. In order to identify Smad3/4 binding to P450arom promoter II that mediates activin A response in ESCs, polymerase chain reaction (PCR) products of a serial truncated deletion of the P450arom promoter II region between -904 and +87 bp were inserted into the pGL3-basic vector to generate the promoter reporter plasmids. Luciferase reporter plasmids were cotransfected into cells with or without activin A (25 ng/mL). The pGL3 -705/+87 revealed a luciferase activity similar to pGL3 -904/+87, whereas progressive truncation to position -464/+87 and -192/+87, the luciferase activity was significant variation. Chromatin immunoprecipitation assay and Smad4-small interfering RNA (siRNA) testify that Smad3/4 binds to the activin A-responsive aromatase promoter in ESCs. Chromatin immunoprecipitation assay-PCR assay demonstrated anti-Smad3 antibody complexes could interact with the amplified DNA of the activin A-responsive P450arom promoter. Mutations of the binding site (-141/-138 bp, -165/-162 bp) in P450arom promoter II significantly reduced promoter activity of activin A fold-induction to 26% and 28%, respectively. We cotransfected pGL3 -705/+87 with control siRNA and Smad4-siRNA into ESCs in the presence of activin A. Luciferase analysis showed that Smad4-siRNA abolished increased promoter activity of activin A-induced P450arom expression. The effect of activin A on the p-Smad3 accumulation in the cytoplasm and nucleus was significantly abrogated following the pretreatment of ESCs with Smad4-siRNA. In conclusion, activated Smad3 proteins can bind to P450arom promoter -705/+87 bp region, responsive to activin A in ESCs, which can promote P450arom transcription.

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Crossref · 10.1177/1933719116681517

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Authors
Qu, J., Zhu, Y., Wu, X., Zheng, J., Hou, Z., Cui, Y., Mao, Y., & Liu, J.
Affiliations
Jiangsu Province Hospital (CN)
Corresponding author
Hou, Zhen
Venue
Reproductive Sciences
Published
2017-08
Volume
24
Issue
8
Pages
1187-1194
ISSN
  • 1933-7191 (ISSN-L)
  • 1933-7205
OpenAlex
W2560811930
Cited by
12
References
45
Citations by year
2025: 3 · 2023: 3 · 2020: 2 · 2019: 2 · 2018: 2
FWCI
1.77
Type
journal article
Language
en
Open access
closed
Locations
  • Reproductive Sciences (publishedVersion)
  • PubMed (publishedVersion)
Topics
  • Health Sciences › Medicine › Reproductive Medicine › Endometriosis Research and Treatment
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › TGF-β signaling in diseases
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
Keywords
Smad3/4, P450arom, aromatase, endometriosis, activin A, endometrial stromal cells, transcriptional regulation, SMAD4, Smad3, activin type I receptor, Smad3 phosphorylation
MeSH terms
Aromatase, Endometriosis, Endometrium, Female, Humans, Promoter Regions, Genetic, Transcription, Genetic, Stromal Cells, Activins, Smad3 Protein, Smad4 Protein

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