In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions

Autor: Renaud Vincent, Andrew Williams, Dalibor Breznan, Julie S. O’Brien, Ngoc Q. Vuong, Patrick Goegan, Premkumari Kumarathasan, Subramanian Karthikeyan
Jazyk: angličtina
Rok vydání: 2017
Předmět:
0301 basic medicine
Proteomics
Vascular Endothelial Growth Factor A
Toxicoproteomics
Health
Toxicology and Mutagenesis
Cytotoxicity
010501 environmental sciences
Toxicology
01 natural sciences
A549
Tandem Mass Spectrometry
Electrophoresis
Gel
Two-Dimensional
Soluble fraction
Lung
Chemokine CCL2
Gel electrophoresis
Chemistry
General Medicine
Inflammatory Response Pathway
Mass spectrometry (MS)
Biochemistry
Environmental chemistry
Toxicity
Inflammation Mediators
Two-dimensional gel electrophoresis (2D–GE)
Signal Transduction
Cell Survival
lcsh:Industrial hygiene. Industrial welfare
Risk Assessment
03 medical and health sciences
lcsh:RA1190-1270
Humans
Secretion
lcsh:Toxicology. Poisons
0105 earth and related environmental sciences
Particulate matter (PM)
A549 cell
Dose-Response Relationship
Drug
Cell growth
Research
Interleukin-8
Water
In vitro
030104 developmental biology
Solubility
A549 Cells
Spectrometry
Mass
Matrix-Assisted Laser Desorption-Ionization
Solvents
Particulate Matter
Insoluble fraction
EHC-93
lcsh:HD7260-7780.8
Zdroj: Particle and Fibre Toxicology
Particle and Fibre Toxicology, Vol 14, Iss 1, Pp 1-19 (2017)
ISSN: 1743-8977
Popis: Background Toxicity of airborne particulate matter (PM) is difficult to assess because PM composition is complex and variable due to source contribution and atmospheric transformation. In this study, we used an in vitro toxicoproteomic approach to identify the toxicity mechanisms associated with different subfractions of Ottawa urban dust (EHC-93). Methods A549 human lung epithelial cells were exposed to 0, 60, 140 and 200 μg/cm2 doses of EHC-93 (total), its insoluble and soluble fractions for 24 h. Multiple cytotoxicity assays and proteomic analyses were used to assess particle toxicity in the exposed cells. Results The cytotoxicity data based on cellular ATP, BrdU incorporation and LDH leakage indicated that the insoluble, but not the soluble, fraction is responsible for the toxicity of EHC-93 in A549 cells. Two-dimensional gel electrophoresis results revealed that the expressions of 206 protein spots were significantly altered after particle exposures, where 154 were identified by MALDI-TOF-TOF-MS/MS. The results from cytotoxicity assays and proteomic analyses converged to a similar finding that the effects of the total and insoluble fraction may be alike, but their effects were distinguishable, and their effects were significantly different from the soluble fraction. Furthermore, the toxic potency of EHC-93 total is not equal to the sum of its insoluble and soluble fractions, implying inter-component interactions between insoluble and soluble materials resulting in synergistic or antagonistic cytotoxic effects. Pathway analysis based on the low toxicity dose (60 μg/cm2) indicated that the two subfractions can alter the expression of those proteins involved in pathways including cell death, cell proliferation and inflammatory response in a distinguishable manner. For example, the insoluble and soluble fractions differentially affected the secretion of pro-inflammatory cytokines such as MCP-1 and IL-8 and distinctly altered the expression of those proteins (e.g., TREM1, PDIA3 and ENO1) involved in an inflammatory response pathway in A549 cells. Conclusions This study demonstrated the impact of different fractions of urban air particles constituted of various chemical species on different mechanistic pathways and thus on cytotoxicity effects. In vitro toxicoproteomics can be a valuable tool in mapping these differences in air pollutant exposure-related toxicity mechanisms. Electronic supplementary material The online version of this article (10.1186/s12989-017-0220-6) contains supplementary material, which is available to authorized users.
Databáze: OpenAIRE
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