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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Published in:Molecular and cellular biochemistry 2019-12, Vol.462 (1-2), p.11-23
Main Authors: Morrone, Maurilio da Silva, Somensi, Nauana, Franz, Lucas, Ramos, Vitor de Miranda, Gasparotto, Juciano, da Rosa, Helen Taís, Sartori, Marcelo, Figueiró, Fabrício, Gelain, Daniel Pens, Zanotto-Filho, Alfeu, Moreira, José Cláudio Fonseca
Format: Article
Language:English
Subjects:
Animals
Biochemistry
Biomarkers - metabolism
Biomedical and Life Sciences
BRCA1 protein
BRCA1 Protein - metabolism
Cardiology
Cell activation
Cell cycle
Cell Cycle - drug effects
Cell differentiation
Cell Differentiation - drug effects
Cell Line, Tumor
Cell Polarity - drug effects
Cell Shape - drug effects
Cell survival
Cell Survival - drug effects
Conditioning
Culture Media, Conditioned - pharmacology
Cytokines
Deoxyribonucleic acid
Depletion
Differentiation
DNA
DNA Damage
DNA repair
Gene regulation
Glioma cells
Immunofluorescence
Inflammation
Inflammation - metabolism
Inflammation - pathology
Interleukin 10
Interleukin 4
Interleukin 6
Life Sciences
Lipopolysaccharides
Macrophage Activation - drug effects
Macrophages
Macrophages - drug effects
Macrophages - metabolism
Macrophages - pathology
Markers
Matrix Metalloproteinases - metabolism
Medical Biochemistry
Mice
Molecular docking
NF-kappa B - metabolism
NF-κB protein
Oncology
Phagocytes
Phagocytosis
Phagocytosis - drug effects
Phenotypes
Rats
RAW 264.7 Cells
Reporter gene
RNA, Small Interfering - metabolism
siRNA
Transcription
Tumor cells
Tumor necrosis factor-α
Tumors
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Summary: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.
ISSN:0300-8177
1573-4919
DOI:10.1007/s11010-019-03605-9