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Breast Cancer Research
Published in: Breast Cancer Research 6/2012

Open Access 01-12-2012 | Research article

Maintenance of S-nitrosothiol homeostasis plays an important role in growth suppression of estrogen receptor-positive breast tumors

Authors: Amanda Cañas, Laura M López-Sánchez, Araceli Valverde-Estepa, Vanessa Hernández, Elena Fuentes, Juan R Muñoz-Castañeda, Chary López-Pedrera, Juan R De La Haba-Rodríguez, Enrique Aranda, Antonio Rodríguez-Ariza

Published in: Breast Cancer Research | Issue 6/2012

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Abstract

Introduction

Protein denitrosylation by thioredoxin reductase (TrxR) is key for maintaining S-nitrosothiol (SNO) homeostasis, although its role in tumor progression is unknown. Therefore, the present study aimed to assess the role of altered SNO homeostasis in breast cancer cells.

Methods

The impairment of SNO homeostasis in breast cancer cells was achieved with the highly specific TrxR inhibitor auranofin and/or exposure to S-nitroso-L-cysteine. S-nitrosylated proteins were detected using the biotin switch assay. Estrogen receptor (ER) alpha knockdown was achieved using RNA silencing technologies and subcellular localization of ERα was analyzed by confocal microscopy. The Oncomine database was explored for TrxR1 (TXNRD1) expression in breast tumors and TrxR1, ER and p53 expression was analyzed by immunohistochemistry in a panel of breast tumors.

Results

The impairment of SNO homeostasis enhanced cell proliferation and survival of ER+ MCF-7 cells, but not of MDA-MB-231 (ER-, mut p53) or BT-474 (ER+, mut p53) cells. This enhanced cell growth and survival was associated with Akt, Erk1/2 phosphorylation, and augmented cyclin D1 expression and was abolished by the ER antagonist fulvestrant or the p53 specific inhibitor pifithrin-α. The specific silencing of ERα expression in MCF-7 cells also abrogated the growth effect of TrxR inhibition. Estrogenic deprivation in MCF-7 cells potentiated the pro-proliferative effect of impaired SNO homeostasis. Moreover, the subcellular distribution of ERα was altered, with a predominant nuclear localization associated with phosphorylation at Thr311 in those cells with impaired SNO homeostasis. The impairment of SNO homeostasis also expanded a cancer stem cell-like subpopulation in MCF-7 cells, as indicated by the increase of percentage of CD44+ cells and the augmented capability to form mammospheres in vitro. Notably, ER+ status in breast tumors was significantly associated with lower TXNDR1 mRNA expression and immunohistochemical studies confirmed this association, particularly when p53 abnormalities were absent.

Conclusion

The ER status in breast cancer may dictate tumor response to different nitrosative environments. Impairment of SNO homeostasis confers survival advantages to ER+ breast tumors, and these molecular mechanisms may also participate in the development of resistance against hormonal therapies that arise in this type of mammary tumors.
Appendix
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Metadata
Title
Maintenance of S-nitrosothiol homeostasis plays an important role in growth suppression of estrogen receptor-positive breast tumors
Authors
Amanda Cañas
Laura M López-Sánchez
Araceli Valverde-Estepa
Vanessa Hernández
Elena Fuentes
Juan R Muñoz-Castañeda
Chary López-Pedrera
Juan R De La Haba-Rodríguez
Enrique Aranda
Antonio Rodríguez-Ariza
Publication date
01-12-2012
Publisher
BioMed Central
Published in
Breast Cancer Research / Issue 6/2012
Electronic ISSN: 1465-542X
DOI
https://doi.org/10.1186/bcr3366

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