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A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide

ABSTRACT: The exposure of HeLa cells to interleukin-1 alpha (IL-1α) in the presence of cycloheximide (CHX) leads to the release of active tumor necrosis factor alpha (TNF-α), eliciting cytocidal effect on these cells. A mass spectrometry (MS)-based analysis of the qualitative proteomic profiles of t...

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Autores principales: Macur, Katarzyna, Grzenkowicz-Wydra, Jolanta, Konieczna, Lucyna, Bigda, Jacek, Temporini, Caterina, Tengattini, Sara, Bączek, Tomasz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5780535/
https://www.ncbi.nlm.nih.gov/pubmed/29398714
http://dx.doi.org/10.1007/s10337-017-3382-3
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author Macur, Katarzyna
Grzenkowicz-Wydra, Jolanta
Konieczna, Lucyna
Bigda, Jacek
Temporini, Caterina
Tengattini, Sara
Bączek, Tomasz
author_facet Macur, Katarzyna
Grzenkowicz-Wydra, Jolanta
Konieczna, Lucyna
Bigda, Jacek
Temporini, Caterina
Tengattini, Sara
Bączek, Tomasz
author_sort Macur, Katarzyna
collection PubMed
description ABSTRACT: The exposure of HeLa cells to interleukin-1 alpha (IL-1α) in the presence of cycloheximide (CHX) leads to the release of active tumor necrosis factor alpha (TNF-α), eliciting cytocidal effect on these cells. A mass spectrometry (MS)-based analysis of the qualitative proteomic profiles of the HeLa cells treated only with IL-1α, CHX or simultaneously with IL-1α and CHX, in comparison to an untreated control, enabled to distinguish protein candidates possibly involved in this process. Among them protein disulphide isomerase (PDI) seemed to be particularly interesting for further research. Therefore, we focused on quantitative changes of PDI levels in HeLa cells subjected to IL-1α and CHX. Enzyme-linked immunosorbent assay (ELISA) was employed for determination of PDI concentrations in the investigated, differently treated HeLa cells. The obtained results confirmed up-regulation of PDI only in the cells stimulated with IL-1α alone. In contrary, the PDI levels in HeLa cells exposed to both IL-1α and CHX, where apoptotic process was intensive, did not increase significantly. Finally, we discuss how different expression levels of PDI together with other proteins, which were detected in this study, may influence the induction of cytotoxic effect and modulate sensitivity to cytotoxic action of IL1. GRAPHICAL ABSTRACT: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10337-017-3382-3) contains supplementary material, which is available to authorized users.
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spelling pubmed-57805352018-02-01 A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide Macur, Katarzyna Grzenkowicz-Wydra, Jolanta Konieczna, Lucyna Bigda, Jacek Temporini, Caterina Tengattini, Sara Bączek, Tomasz Chromatographia Original ABSTRACT: The exposure of HeLa cells to interleukin-1 alpha (IL-1α) in the presence of cycloheximide (CHX) leads to the release of active tumor necrosis factor alpha (TNF-α), eliciting cytocidal effect on these cells. A mass spectrometry (MS)-based analysis of the qualitative proteomic profiles of the HeLa cells treated only with IL-1α, CHX or simultaneously with IL-1α and CHX, in comparison to an untreated control, enabled to distinguish protein candidates possibly involved in this process. Among them protein disulphide isomerase (PDI) seemed to be particularly interesting for further research. Therefore, we focused on quantitative changes of PDI levels in HeLa cells subjected to IL-1α and CHX. Enzyme-linked immunosorbent assay (ELISA) was employed for determination of PDI concentrations in the investigated, differently treated HeLa cells. The obtained results confirmed up-regulation of PDI only in the cells stimulated with IL-1α alone. In contrary, the PDI levels in HeLa cells exposed to both IL-1α and CHX, where apoptotic process was intensive, did not increase significantly. Finally, we discuss how different expression levels of PDI together with other proteins, which were detected in this study, may influence the induction of cytotoxic effect and modulate sensitivity to cytotoxic action of IL1. GRAPHICAL ABSTRACT: [Image: see text] ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s10337-017-3382-3) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2017-08-30 2018 /pmc/articles/PMC5780535/ /pubmed/29398714 http://dx.doi.org/10.1007/s10337-017-3382-3 Text en © The Author(s) 2017 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original
Macur, Katarzyna
Grzenkowicz-Wydra, Jolanta
Konieczna, Lucyna
Bigda, Jacek
Temporini, Caterina
Tengattini, Sara
Bączek, Tomasz
A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide
title A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide
title_full A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide
title_fullStr A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide
title_full_unstemmed A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide
title_short A Proteomic-Based Approach to Study the Mechanism of Cytotoxicity Induced by Interleukin-1α and Cycloheximide
title_sort proteomic-based approach to study the mechanism of cytotoxicity induced by interleukin-1α and cycloheximide
topic Original
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5780535/
https://www.ncbi.nlm.nih.gov/pubmed/29398714
http://dx.doi.org/10.1007/s10337-017-3382-3
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