Smad3/4 Binding to Promoter II of P450arom So As to Regulate Aromatase Expression in Endometriosis
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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TGF-β1/Smad3 signaling pathway is involved in the transcriptional regulation of aromatase in several cell types, and particularly in endometriotic stromal cells.
“Previous studies have reported that the TGF-β1/Smad3 signaling pathway is involved in the transcriptional regulation of aromatase in several cell types56–58, and particularly in endometriotic stromal cells59.”
From Platelets induce increased estrogen production through NF-κB and TGF-β1 signaling pathways in endometriotic stromal cells (2020) · cites this paper as [59]
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Pathogenesis of endometriosis: The genetic/epigenetic theory
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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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