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Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin

The Sall2 transcription factor is deregulated in several cancers; however, little is known about its cellular functions, including its target genes. Recently, we demonstrated that p53 directly regulates Sall2 expression under genotoxic stress. Here, we investigated the role of Sall2 in the context o...

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Autores principales: Escobar, D, Hepp, M I, Farkas, C, Campos, T, Sodir, N M, Morales, M, Álvarez, C I, Swigart, L, Evan, G I, Gutiérrez, J L, Nishinakamura, R, Castro, A F, Pincheira, R
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4650718/
https://www.ncbi.nlm.nih.gov/pubmed/26181197
http://dx.doi.org/10.1038/cddis.2015.165
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author Escobar, D
Hepp, M I
Farkas, C
Campos, T
Sodir, N M
Morales, M
Álvarez, C I
Swigart, L
Evan, G I
Gutiérrez, J L
Nishinakamura, R
Castro, A F
Pincheira, R
author_facet Escobar, D
Hepp, M I
Farkas, C
Campos, T
Sodir, N M
Morales, M
Álvarez, C I
Swigart, L
Evan, G I
Gutiérrez, J L
Nishinakamura, R
Castro, A F
Pincheira, R
author_sort Escobar, D
collection PubMed
description The Sall2 transcription factor is deregulated in several cancers; however, little is known about its cellular functions, including its target genes. Recently, we demonstrated that p53 directly regulates Sall2 expression under genotoxic stress. Here, we investigated the role of Sall2 in the context of cellular response to genotoxic stress. In addition, we further examined the Sall2-p53 relationship during genotoxic stress in primary mouse embryo fibroblasts (MEFs), which are derived from Sall2 knockout mice separately, or in combination with the p53ERTAM knock-in mice. We found that the levels of Sall2 mRNA and protein are dynamically modulated in response to doxorubicin. At early times of stress, Sall2 is downregulated, but increases under extension of the stress in a p53-independent manner. Based on caspase-3/7 activities, expression of cleaved poly (ADP-ribose) polymerase, expression of cleaved caspase-3 and induction of proapoptotic proteins, Sall2 expression was correlated with cellular apoptosis. Consequently, Sall2(−/−) MEFs have decreased apoptosis, which relates with increased cell viability in response to doxorubicin. Importantly, Sall2 was required for apoptosis even in the presence of fully activated p53. Searching for putative Sall2 targets that could mediate its role in apoptosis, we identified proapoptotic NOXA/PMAIP1 (phorbol-12-myristate-13-acetate-induced protein 1). We demonstrated that Sall2 positively regulates Noxa promoter activity. Conserved putative Sall2-binding sites at the NOXA promoter were validated in vitro by electrophoretic mobility shift assay and in vivo by ChIP experiments, identifying NOXA as a novel Sall2 target. In agreement, induction of Noxa protein and mRNA in response to doxorubicin was significantly decreased in Sall2(−/−) MEFs. In addition, studies in leukemia Jurkat T cells support the existence of the Sall2/Noxa axis, and the significance of this axis on the apoptotic response to doxorubicin in cancer cells. Our study highlights the relevance of Sall2 in the apoptotic response to extended genotoxic stress, which is important for understanding its role in normal physiology and disease.
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spelling pubmed-46507182015-12-02 Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin Escobar, D Hepp, M I Farkas, C Campos, T Sodir, N M Morales, M Álvarez, C I Swigart, L Evan, G I Gutiérrez, J L Nishinakamura, R Castro, A F Pincheira, R Cell Death Dis Original Article The Sall2 transcription factor is deregulated in several cancers; however, little is known about its cellular functions, including its target genes. Recently, we demonstrated that p53 directly regulates Sall2 expression under genotoxic stress. Here, we investigated the role of Sall2 in the context of cellular response to genotoxic stress. In addition, we further examined the Sall2-p53 relationship during genotoxic stress in primary mouse embryo fibroblasts (MEFs), which are derived from Sall2 knockout mice separately, or in combination with the p53ERTAM knock-in mice. We found that the levels of Sall2 mRNA and protein are dynamically modulated in response to doxorubicin. At early times of stress, Sall2 is downregulated, but increases under extension of the stress in a p53-independent manner. Based on caspase-3/7 activities, expression of cleaved poly (ADP-ribose) polymerase, expression of cleaved caspase-3 and induction of proapoptotic proteins, Sall2 expression was correlated with cellular apoptosis. Consequently, Sall2(−/−) MEFs have decreased apoptosis, which relates with increased cell viability in response to doxorubicin. Importantly, Sall2 was required for apoptosis even in the presence of fully activated p53. Searching for putative Sall2 targets that could mediate its role in apoptosis, we identified proapoptotic NOXA/PMAIP1 (phorbol-12-myristate-13-acetate-induced protein 1). We demonstrated that Sall2 positively regulates Noxa promoter activity. Conserved putative Sall2-binding sites at the NOXA promoter were validated in vitro by electrophoretic mobility shift assay and in vivo by ChIP experiments, identifying NOXA as a novel Sall2 target. In agreement, induction of Noxa protein and mRNA in response to doxorubicin was significantly decreased in Sall2(−/−) MEFs. In addition, studies in leukemia Jurkat T cells support the existence of the Sall2/Noxa axis, and the significance of this axis on the apoptotic response to doxorubicin in cancer cells. Our study highlights the relevance of Sall2 in the apoptotic response to extended genotoxic stress, which is important for understanding its role in normal physiology and disease. Nature Publishing Group 2015-07 2015-07-16 /pmc/articles/PMC4650718/ /pubmed/26181197 http://dx.doi.org/10.1038/cddis.2015.165 Text en Copyright © 2015 Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ Cell Death and Disease is an open-access journal published by Nature Publishing Group. This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Original Article
Escobar, D
Hepp, M I
Farkas, C
Campos, T
Sodir, N M
Morales, M
Álvarez, C I
Swigart, L
Evan, G I
Gutiérrez, J L
Nishinakamura, R
Castro, A F
Pincheira, R
Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin
title Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin
title_full Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin
title_fullStr Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin
title_full_unstemmed Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin
title_short Sall2 is required for proapoptotic Noxa expression and genotoxic stress-induced apoptosis by doxorubicin
title_sort sall2 is required for proapoptotic noxa expression and genotoxic stress-induced apoptosis by doxorubicin
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4650718/
https://www.ncbi.nlm.nih.gov/pubmed/26181197
http://dx.doi.org/10.1038/cddis.2015.165
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