TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.

There is increasing evidence that soluble factors in inflammatory central nervous system diseases not only regulate the inflammatory process but also directly influence electrophysiological membrane properties of neurons and astrocytes. In this context, the cytokine TNF-α (tumor necrosis factor-α) h...

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Autores principales:	Susana Alvarez, Almudena Blanco, Manuel Fresno, Ma Ángeles Muñoz-Fernández
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2011
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9d74a52905354ad2aa592b4e4a8c6361
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spelling	oai:doaj.org-article:9d74a52905354ad2aa592b4e4a8c63612021-11-18T06:59:45ZTNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.1932-620310.1371/journal.pone.0016100https://doaj.org/article/9d74a52905354ad2aa592b4e4a8c63612011-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/21298033/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203There is increasing evidence that soluble factors in inflammatory central nervous system diseases not only regulate the inflammatory process but also directly influence electrophysiological membrane properties of neurons and astrocytes. In this context, the cytokine TNF-α (tumor necrosis factor-α) has complex injury promoting, as well as protective, effects on neuronal viability. Up-regulated TNF-α expression has also been found in various neurodegenerative diseases such as cerebral malaria, AIDS dementia, Alzheimer's disease, multiple sclerosis, and stroke, suggesting a potential pathogenic role of TNF-α in these diseases as well. We used the neuroblastoma cells SK-N-MC. Transcriptional activity was measured using luciferase reporter gene assays by using lipofectin. We performed cotransfection experiments of NFAT (nuclear factor of activated T cells) promoter constructed with a dominant negative version of NFAT (dn-NFAT). Cell death was performed by MTT (3-(4,5-dimethylthiazol-2-yl)5,5-diphenyltetrazolium bromide) and TUNEL assays. NFAT translocation was confirmed by Western blot. Involvement of NFAT in cell death was assessed by using VIVIT. P53, Fas-L, caspase-3, and caspase-9 expressions were carried out by Western blot. The mechanisms involved in TNF-α-induced cell death were assessed by using microarray analysis. TNF-α causes neuronal cell death in the absence of glia. TNF-α treatment results in nuclear translocation of NFAT through activation of calcineurin in a Ca(2+) independent manner. We demonstrated the involvement of FasL/Fas, cytochrome c, and caspase-9 but the lack of caspase-3 activation. NB cell death was absolutely reverted in the presence of VIVIT, and partially diminished by anti-Fas treatment. These data demonstrate that TNF-α promotes FasL expression through NFAT activation in neuroblastoma cells and this event leads to increased apoptosis through independent caspase-3 activation.Susana AlvarezAlmudena BlancoManuel FresnoMa Ángeles Muñoz-FernándezPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 6, Iss 1, p e16100 (2011)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Susana Alvarez Almudena Blanco Manuel Fresno Ma Ángeles Muñoz-Fernández TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.
description	There is increasing evidence that soluble factors in inflammatory central nervous system diseases not only regulate the inflammatory process but also directly influence electrophysiological membrane properties of neurons and astrocytes. In this context, the cytokine TNF-α (tumor necrosis factor-α) has complex injury promoting, as well as protective, effects on neuronal viability. Up-regulated TNF-α expression has also been found in various neurodegenerative diseases such as cerebral malaria, AIDS dementia, Alzheimer's disease, multiple sclerosis, and stroke, suggesting a potential pathogenic role of TNF-α in these diseases as well. We used the neuroblastoma cells SK-N-MC. Transcriptional activity was measured using luciferase reporter gene assays by using lipofectin. We performed cotransfection experiments of NFAT (nuclear factor of activated T cells) promoter constructed with a dominant negative version of NFAT (dn-NFAT). Cell death was performed by MTT (3-(4,5-dimethylthiazol-2-yl)5,5-diphenyltetrazolium bromide) and TUNEL assays. NFAT translocation was confirmed by Western blot. Involvement of NFAT in cell death was assessed by using VIVIT. P53, Fas-L, caspase-3, and caspase-9 expressions were carried out by Western blot. The mechanisms involved in TNF-α-induced cell death were assessed by using microarray analysis. TNF-α causes neuronal cell death in the absence of glia. TNF-α treatment results in nuclear translocation of NFAT through activation of calcineurin in a Ca(2+) independent manner. We demonstrated the involvement of FasL/Fas, cytochrome c, and caspase-9 but the lack of caspase-3 activation. NB cell death was absolutely reverted in the presence of VIVIT, and partially diminished by anti-Fas treatment. These data demonstrate that TNF-α promotes FasL expression through NFAT activation in neuroblastoma cells and this event leads to increased apoptosis through independent caspase-3 activation.
format	article
author	Susana Alvarez Almudena Blanco Manuel Fresno Ma Ángeles Muñoz-Fernández
author_facet	Susana Alvarez Almudena Blanco Manuel Fresno Ma Ángeles Muñoz-Fernández
author_sort	Susana Alvarez
title	TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.
title_short	TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.
title_full	TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.
title_fullStr	TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.
title_full_unstemmed	TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.
title_sort	tnf-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of nfat.
publisher	Public Library of Science (PLoS)
publishDate	2011
url	https://doaj.org/article/9d74a52905354ad2aa592b4e4a8c6361
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TNF-α contributes to caspase-3 independent apoptosis in neuroblastoma cells: role of NFAT.

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