Using SILAC proteomics to investigate the effect of the mycotoxin, alternariol, in the human H295R steroidogenesis model
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Date
2014-11
Journal Title
Journal ISSN
Volume Title
Publisher
Springer Science+Business Media Dordrecht 2014
Abstract
The mycotoxin alternariol (AOH) is an impor-
tant contaminant of fruits and cereal products. The current
study sought to address the effect of a non-toxic AOH
concentration on the proteome of the steroidogenic H295R
cell model. Quantitative proteomics based on stable iso-
tope labeling by amino acids in cell culture (SILAC)
coupled to 1D-SDS-PAGE-LC-MS/MS was applied to
subcellular-enriched protein samples. Gene ontology (GO) and ingenuity pathway analysis (IPA) were further carried out for functional annotation and identification of
protein interaction networks. Furthermore, the effect of
AOH on apoptosis and cell cycle distribution was also
determined by the use of flow cytometry analysis. This
work identified 22 proteins that were regulated significant-
ly. The regulated proteins are those involved in early stages
of steroid biosynthesis (SOAT1, NPC1, and ACBD5) and
C21-steroid hormone metabolism (CYP21A2 and
HSD3B1). In addition, several proteins known to play a
role in cellular assembly, organization, protein synthesis,
and cell cycle were regulated. These findings provide a
new framework for studying the mechanisms by which
AOH modulates steroidogenesis in H295R cell model.
Description
Journal of Cell Biol Toxicol (2014) 30:pp. 361–376
Keywords
Alternariol, Mycotoxins, SILAC, Sterodogenesis, Endocrine disruption, Quantitative proteomics