In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions
Abstract 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...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-10-01
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| Series: | Particle and Fibre Toxicology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s12989-017-0220-6 |
| _version_ | 1818475758822096896 |
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| author | Ngoc Q. Vuong Dalibor Breznan Patrick Goegan Julie S. O’Brien Andrew Williams Subramanian Karthikeyan Premkumari Kumarathasan Renaud Vincent |
| author_facet | Ngoc Q. Vuong Dalibor Breznan Patrick Goegan Julie S. O’Brien Andrew Williams Subramanian Karthikeyan Premkumari Kumarathasan Renaud Vincent |
| author_sort | Ngoc Q. Vuong |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-10T09:16:45Z |
| format | Article |
| id | doaj.art-e6f2b9510afd4826b89d011e9fb5c621 |
| institution | Directory Open Access Journal |
| issn | 1743-8977 |
| language | English |
| last_indexed | 2024-12-10T09:16:45Z |
| publishDate | 2017-10-01 |
| publisher | BMC |
| record_format | Article |
| series | Particle and Fibre Toxicology |
| spelling | doaj.art-e6f2b9510afd4826b89d011e9fb5c6212022-12-22T01:54:51ZengBMCParticle and Fibre Toxicology1743-89772017-10-0114111910.1186/s12989-017-0220-6In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractionsNgoc Q. Vuong0Dalibor Breznan1Patrick Goegan2Julie S. O’Brien3Andrew Williams4Subramanian Karthikeyan5Premkumari Kumarathasan6Renaud Vincent7Inhalation Toxicology Laboratory, Environmental Health Science and Research Bureau, Health CanadaInhalation Toxicology Laboratory, Environmental Health Science and Research Bureau, Health CanadaInhalation Toxicology Laboratory, Environmental Health Science and Research Bureau, Health CanadaInhalation Toxicology Laboratory, Environmental Health Science and Research Bureau, Health CanadaBiostatistics Section, Population Studies Division, Environmental Health Science and Research Bureau, Health CanadaInhalation Toxicology Laboratory, Environmental Health Science and Research Bureau, Health CanadaAnalytical Biochemistry and Proteomics, Environmental Health Science and Research Bureau, Health CanadaInhalation Toxicology Laboratory, Environmental Health Science and Research Bureau, Health CanadaAbstract 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.http://link.springer.com/article/10.1186/s12989-017-0220-6Particulate matter (PM)EHC-93Soluble fractionInsoluble fractionA549Cytotoxicity |
| spellingShingle | Ngoc Q. Vuong Dalibor Breznan Patrick Goegan Julie S. O’Brien Andrew Williams Subramanian Karthikeyan Premkumari Kumarathasan Renaud Vincent In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions Particle and Fibre Toxicology Particulate matter (PM) EHC-93 Soluble fraction Insoluble fraction A549 Cytotoxicity |
| title | In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions |
| title_full | In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions |
| title_fullStr | In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions |
| title_full_unstemmed | In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions |
| title_short | In vitro toxicoproteomic analysis of A549 human lung epithelial cells exposed to urban air particulate matter and its water-soluble and insoluble fractions |
| title_sort | in vitro toxicoproteomic analysis of a549 human lung epithelial cells exposed to urban air particulate matter and its water soluble and insoluble fractions |
| topic | Particulate matter (PM) EHC-93 Soluble fraction Insoluble fraction A549 Cytotoxicity |
| url | http://link.springer.com/article/10.1186/s12989-017-0220-6 |
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