BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism

BRCA-1 is a nuclear protein involved in DNA repair, transcriptional regulation, and cell cycle control. Its involvement in other cellular processes has been described. Here, we aimed to investigate the role of BRCA-1 in macrophages M(LPS), M(IL-4), and tumor cell-induced differentiation. We used siR...

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Autores:
da Silva Morrone, Maurilio
Somensi, Nauana
Franz, Lucas
de Miranda Ramos, Vitor
Gasparotto, Juciano
Taís da Rosa, Helen
Sartori, Marcelo
Figueiró, Fabrício
Gelain, Daniel
ZANOTTO-FILHO, ALFEU
Moreira, José Cláudio Fonseca
Tipo de recurso:
http://purl.org/coar/resource_type/c_816b
Fecha de publicación:
2019
Institución:
Corporación Universidad de la Costa
Repositorio:
REDICUC - Repositorio CUC
Idioma:
eng
OAI Identifier:
oai:repositorio.cuc.edu.co:11323/5657
Acceso en línea:
https://hdl.handle.net/11323/5657
https://repositorio.cuc.edu.co/
Palabra clave:
BRCA-1
Inflammation
M(LPS) and M(IL-4) polarization
Macrophage activation
Rights
openAccess
License
CC0 1.0 Universal
id RCUC2_08a3686156369be44859d416e044bfd8
oai_identifier_str oai:repositorio.cuc.edu.co:11323/5657
network_acronym_str RCUC2
network_name_str REDICUC - Repositorio CUC
repository_id_str
dc.title.spa.fl_str_mv BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
title BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
spellingShingle BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
BRCA-1
Inflammation
M(LPS) and M(IL-4) polarization
Macrophage activation
title_short BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
title_full BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
title_fullStr BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
title_full_unstemmed BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
title_sort BRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism
dc.creator.fl_str_mv da Silva Morrone, Maurilio
Somensi, Nauana
Franz, Lucas
de Miranda Ramos, Vitor
Gasparotto, Juciano
Taís da Rosa, Helen
Sartori, Marcelo
Figueiró, Fabrício
Gelain, Daniel
ZANOTTO-FILHO, ALFEU
Moreira, José Cláudio Fonseca
dc.contributor.author.spa.fl_str_mv da Silva Morrone, Maurilio
Somensi, Nauana
Franz, Lucas
de Miranda Ramos, Vitor
Gasparotto, Juciano
Taís da Rosa, Helen
Sartori, Marcelo
Figueiró, Fabrício
Gelain, Daniel
ZANOTTO-FILHO, ALFEU
Moreira, José Cláudio Fonseca
dc.subject.spa.fl_str_mv BRCA-1
Inflammation
M(LPS) and M(IL-4) polarization
Macrophage activation
topic BRCA-1
Inflammation
M(LPS) and M(IL-4) polarization
Macrophage activation
description BRCA-1 is a nuclear protein involved in DNA repair, transcriptional regulation, and cell cycle control. Its involvement in other cellular processes has been described. Here, we aimed to investigate the role of BRCA-1 in macrophages M(LPS), M(IL-4), and tumor cell-induced differentiation. We used siRNAs to knockdown BRCA-1 in RAW 264.7 macrophages exposed to LPS, IL-4, and C6 glioma cells conditioned medium (CMC6), and evaluated macrophage differentiation markers and functional phagocytic activity as well as DNA damage and cell survival in the presence and absence of BRCA-1. LPS and CMC6, but not by IL-4, increased DNA damage in macrophages, and this effect was more pronounced in BRCA-1-depleted cells, including M(IL-4). BRCA-1 depletion impaired expression of pro-inflammatory cytokines, TNF-α and IL-6, and reduced the phagocytic activity of macrophages in response to LPS. In CMC6-induced differentiation, BRCA-1 knockdown inhibited TNF-α and IL-6 expression which was accompanied by upregulation of the anti-inflammatory markers IL-10 and TGF-β and reduced phagocytosis. In contrast, M(IL-4) phenotype was not affected by BRCA-1 status. Molecular docking predicted that the conserved BRCA-1 domain BRCT can interact with the p65 subunit of NF-κB. Immunofluorescence assays showed that BRCA-1 and p65 co-localize in the nucleus of LPS-treated macrophages and reporter gene assay showed that depletion of BRCA-1 decreased LPS and CMC6-induced NF-κB transactivation. IL-4 had no effect upon NF-κB. Taken together, our findings suggest a role of BRCA-1 in macrophage differentiation and phagocytosis induced by LPS and tumor cells secretoma, but not IL-4, in a mechanism associated with inhibition of NF-κB.
publishDate 2019
dc.date.accessioned.none.fl_str_mv 2019-11-14T20:24:41Z
dc.date.available.none.fl_str_mv 2019-11-14T20:24:41Z
dc.date.issued.none.fl_str_mv 2019-12
dc.type.spa.fl_str_mv Pre-Publicación
dc.type.coar.spa.fl_str_mv http://purl.org/coar/resource_type/c_816b
dc.type.content.spa.fl_str_mv Text
dc.type.driver.spa.fl_str_mv info:eu-repo/semantics/preprint
dc.type.redcol.spa.fl_str_mv http://purl.org/redcol/resource_type/ARTOTR
dc.type.version.spa.fl_str_mv info:eu-repo/semantics/acceptedVersion
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status_str acceptedVersion
dc.identifier.uri.spa.fl_str_mv https://hdl.handle.net/11323/5657
dc.identifier.instname.spa.fl_str_mv Corporación Universidad de la Costa
dc.identifier.reponame.spa.fl_str_mv REDICUC - Repositorio CUC
dc.identifier.repourl.spa.fl_str_mv https://repositorio.cuc.edu.co/
url https://hdl.handle.net/11323/5657
https://repositorio.cuc.edu.co/
identifier_str_mv Corporación Universidad de la Costa
REDICUC - Repositorio CUC
dc.language.iso.none.fl_str_mv eng
language eng
dc.rights.spa.fl_str_mv CC0 1.0 Universal
dc.rights.uri.spa.fl_str_mv http://creativecommons.org/publicdomain/zero/1.0/
dc.rights.accessrights.spa.fl_str_mv info:eu-repo/semantics/openAccess
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rights_invalid_str_mv CC0 1.0 Universal
http://creativecommons.org/publicdomain/zero/1.0/
http://purl.org/coar/access_right/c_abf2
eu_rights_str_mv openAccess
dc.publisher.spa.fl_str_mv Universidad de la Costa
institution Corporación Universidad de la Costa
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spelling da Silva Morrone, MaurilioSomensi, NauanaFranz, Lucasde Miranda Ramos, VitorGasparotto, JucianoTaís da Rosa, HelenSartori, MarceloFigueiró, FabrícioGelain, DanielZANOTTO-FILHO, ALFEUMoreira, José Cláudio Fonseca2019-11-14T20:24:41Z2019-11-14T20:24:41Z2019-12https://hdl.handle.net/11323/5657Corporación Universidad de la CostaREDICUC - Repositorio CUChttps://repositorio.cuc.edu.co/BRCA-1 is a nuclear protein involved in DNA repair, transcriptional regulation, and cell cycle control. Its involvement in other cellular processes has been described. Here, we aimed to investigate the role of BRCA-1 in macrophages M(LPS), M(IL-4), and tumor cell-induced differentiation. We used siRNAs to knockdown BRCA-1 in RAW 264.7 macrophages exposed to LPS, IL-4, and C6 glioma cells conditioned medium (CMC6), and evaluated macrophage differentiation markers and functional phagocytic activity as well as DNA damage and cell survival in the presence and absence of BRCA-1. LPS and CMC6, but not by IL-4, increased DNA damage in macrophages, and this effect was more pronounced in BRCA-1-depleted cells, including M(IL-4). BRCA-1 depletion impaired expression of pro-inflammatory cytokines, TNF-α and IL-6, and reduced the phagocytic activity of macrophages in response to LPS. In CMC6-induced differentiation, BRCA-1 knockdown inhibited TNF-α and IL-6 expression which was accompanied by upregulation of the anti-inflammatory markers IL-10 and TGF-β and reduced phagocytosis. In contrast, M(IL-4) phenotype was not affected by BRCA-1 status. Molecular docking predicted that the conserved BRCA-1 domain BRCT can interact with the p65 subunit of NF-κB. Immunofluorescence assays showed that BRCA-1 and p65 co-localize in the nucleus of LPS-treated macrophages and reporter gene assay showed that depletion of BRCA-1 decreased LPS and CMC6-induced NF-κB transactivation. IL-4 had no effect upon NF-κB. Taken together, our findings suggest a role of BRCA-1 in macrophage differentiation and phagocytosis induced by LPS and tumor cells secretoma, but not IL-4, in a mechanism associated with inhibition of NF-κB.da Silva Morrone, Maurilio-will be generated-orcid-0000-0001-8144-0347-600Somensi, Nauana-will be generated-orcid-0000-0003-0440-7249-600Franz, Lucasde Miranda Ramos, Vitor-will be generated-orcid-0000-0001-7004-0823-600Gasparotto, Juciano-will be generated-orcid-0000-0003-2545-7288-600Taís da Rosa, HelenSartori, Marcelo-will be generated-orcid-0000-0003-4999-3122-600Figueiró, Fabrício-will be generated-orcid-0000-0003-0899-6407-600Gelain, Daniel-will be generated-orcid-0000-0001-5254-0509-600ZANOTTO-FILHO, ALFEU-will be generated-orcid-0000-0002-4188-3095-600Moreira, José Cláudio Fonseca-will be generated-orcid-0000-0002-0619-4913-600engUniversidad de la CostaCC0 1.0 Universalhttp://creativecommons.org/publicdomain/zero/1.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2BRCA-1InflammationM(LPS) and M(IL-4) polarizationMacrophage activationBRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanismPre-Publicaciónhttp://purl.org/coar/resource_type/c_816bTextinfo:eu-repo/semantics/preprinthttp://purl.org/redcol/resource_type/ARTOTRinfo:eu-repo/semantics/acceptedVersionPublicationORIGINALBRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism.pdfBRCA-1 depletion impairs pro-inflammatory polarization and activation of RAW 264.7 macrophages in a NF-κB-dependent mechanism.pdfapplication/pdf84775https://repositorio.cuc.edu.co/bitstreams/31297a7f-996a-40f1-88c3-f3536ef5e886/download56b592e650a427016d0d878709adfff7MD51CC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; 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