A proteomic-based approach to study the mechanism of cytotoxicity induced by interleukin-1α and cycloheximide

Katarzyna Macur , Jolanta Grzenkowicz-Wydra , Lucyna Konieczna , Jacek Bigda , Caterina Temporini , Sara Tengattini , Tomasz Bączek

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.
Author Katarzyna Macur (IFB / ZLS)
Katarzyna Macur,,
- Zespół Laboratoriów Specjalistycznych MWB UG i GUMed
, Jolanta Grzenkowicz-Wydra
Jolanta Grzenkowicz-Wydra,,
-
, Lucyna Konieczna
Lucyna Konieczna,,
-
, Jacek Bigda (IFB / DMedB)
Jacek Bigda,,
- Department of Medical Biotechnology
, Caterina Temporini
Caterina Temporini,,
-
, Sara Tengattini
Sara Tengattini,,
-
, Tomasz Bączek
Tomasz Bączek,,
-
Journal seriesChromatographia, ISSN 0009-5893, (A 20 pkt)
Issue year2018
Vol82
No1
Pages47-56
Publication size in sheets0.5
Keywords in EnglishHPLC–MS/MS, Protein disulphide isomerase (PDI), Interleukin-1α (IL-1α), cycloheximide, cytotoxicity, HeLa proteome
DOIDOI:10.1007/s10337-017-3382-3
URL https://link.springer.com/content/pdf/10.1007%2Fs10337-017-3382-3.pdf
Languageen angielski
LicenseOther; published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)20
ScoreMinisterial score = 20.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 20.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 1.402 (2) - 2016=1.302 (5)
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