Abstract
The mycotoxin alternariol (AOH) is an important 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 isotope 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 significantly. 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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Adaptor Proteins, Signal Transducing / genetics
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Adaptor Proteins, Signal Transducing / metabolism
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Adrenal Cortex / drug effects*
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Adrenal Cortex / metabolism
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Adrenal Cortex / pathology
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Apoptosis / drug effects
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Carrier Proteins / genetics
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Carrier Proteins / metabolism
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Cell Cycle / drug effects
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Cell Line, Tumor
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Gene Expression Regulation
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Humans
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Intracellular Signaling Peptides and Proteins
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Isotope Labeling
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Lactones / pharmacology*
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Membrane Glycoproteins / genetics
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Membrane Glycoproteins / metabolism
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Membrane Proteins / genetics
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Membrane Proteins / metabolism
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Metabolic Networks and Pathways / drug effects
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Molecular Sequence Annotation
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Multienzyme Complexes / genetics
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Multienzyme Complexes / metabolism
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Mycotoxins / pharmacology*
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Niemann-Pick C1 Protein
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Progesterone Reductase / genetics
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Progesterone Reductase / metabolism
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Protein Interaction Mapping
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Protein Interaction Maps / drug effects*
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Protein Interaction Maps / genetics
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Proteome / genetics*
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Proteome / metabolism
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Proteomics / methods
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Steroid 21-Hydroxylase / genetics
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Steroid 21-Hydroxylase / metabolism
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Steroid Isomerases / genetics
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Steroid Isomerases / metabolism
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Steroids / biosynthesis*
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Sterol O-Acyltransferase / genetics
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Sterol O-Acyltransferase / metabolism
Substances
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3 beta-hydroxysteroid oxidoreductase-delta(5) 3-ketosteroid isomerase
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ACBD5 protein, human
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Adaptor Proteins, Signal Transducing
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Carrier Proteins
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Intracellular Signaling Peptides and Proteins
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Lactones
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Membrane Glycoproteins
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Membrane Proteins
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Multienzyme Complexes
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Mycotoxins
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NPC1 protein, human
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Niemann-Pick C1 Protein
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Proteome
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Steroids
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Progesterone Reductase
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CYP21A2 protein, human
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Steroid 21-Hydroxylase
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Sterol O-Acyltransferase
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sterol O-acyltransferase 1
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Steroid Isomerases
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alternariol