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Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects

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Resumo:Polycyclic Aromatic Hydrocarbons (PAHs) are persistent pollutants normally found in the environment as complex mixtures. Although several individual PAHs are classified as mutagenic and carcinogenic pollutants, the interaction effects between compounds in a mixture may trigger different toxicological mechanisms and, consequently, yield different effects to organisms which are not accounted for in risk assessment guidelines. Given the ubiquity of PAHs, understanding the mechanistic features of their mixtures is a pressing research need. Therefore, the present work aimed to disclose the interaction effects of three PAHs with different carcinogenic potential and chemical structure, in primary hepatocyte cells of gilt-headed seabreams (Sparus aurata). Hepatocytes were exposed to Phenanthrene (Phe), Benzo[a]pyrene (B[a]P) and Benzo[b]fluoranthene (B[b]F) and their mixtures at different proportions and several cellular responses were analyzed: cellular viability, CYP1A1 activity (EROD assay) and protein expression level (Western blot); transcript (mRNA) levels of CYP1A1, EPXH1 and GST-3 (qRT-PCR); genotoxic effects (DNA strand breakage) by the Comet assay. Results show that B[a]P induced CYP1A1 gene and protein expression increasing its activity and, therefore, increasing the production of metabolites that trigger genotoxic DNA damage (%). Most importantly, mixtures containing Phe and B[a]P increased even further CYP1A1 mRNA levels and DNA damage (up to 70 %) which suggests that, although Phe is considered a non-carcinogenic PAH, it potentiates CYP1A1 synthesis induced by B[a]P, increasing its genotoxicity. These findings indicate that the upregulation of CYP1A1 by carcinogenic PAHs will not weaken even when in mixtures with non-carcinogenic PAHs. On contrary, non-carcinogenic PAHs may potentiate the genotoxic effect of carcinogenic PAH and therefore mixture composition should be taken in account when assessing PAH toxicity. In fact, our results point to the need of redefining Environmental Risk Assessment protocols for mixtures of carcinogenic pollutants.
Autores principais:Bramatti, Isabella
Outros Autores:Matos, Beatriz; Figueiredo, Neusa; Pousão-Ferreira, Pedro; Branco, Vasco; Martins, Marta
Assunto:CYP1A1 DNA strand breakage Phenanthrene Benzo[a]pyrene Benzo[b]fluoranthene
Ano:2023
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:Universidade de Lisboa
Idioma:inglês
Origem:Repositório da Universidade de Lisboa
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author Bramatti, Isabella
author2 Matos, Beatriz
Figueiredo, Neusa
Pousão-Ferreira, Pedro
Branco, Vasco
Martins, Marta
author2_role author
author
author
author
author
author_facet Bramatti, Isabella
Matos, Beatriz
Figueiredo, Neusa
Pousão-Ferreira, Pedro
Branco, Vasco
Martins, Marta
author_role author
contributor_name_str_mv Repositório Científico de Acesso Aberto da ULisboa
country_str PT
creators_json_txt [{\"Person.name\":\"Bramatti, Isabella\"},{\"Person.name\":\"Matos, Beatriz\",\"Person.identifier.orcid\":\"0000-0001-5923-5849\"},{\"Person.name\":\"Figueiredo, Neusa\",\"Person.identifier.orcid\":\"0000-0003-0062-1117\"},{\"Person.name\":\"Pousão-Ferreira, Pedro\",\"Person.identifier.orcid\":\"0000-0001-6746-764X\"},{\"Person.name\":\"Branco, Vasco\",\"Person.identifier.orcid\":\"0000-0002-0466-4934\"},{\"Person.name\":\"Martins, Marta\",\"Person.identifier.orcid\":\"0000-0002-3818-6256\"}]
datacite.contributors.contributor.contributorName.fl_str_mv Repositório Científico de Acesso Aberto da ULisboa
datacite.creators.creator.creatorName.fl_str_mv Bramatti, Isabella
Matos, Beatriz
Figueiredo, Neusa
Pousão-Ferreira, Pedro
Branco, Vasco
Martins, Marta
datacite.date.Accepted.fl_str_mv 2023-01-01T00:00:00Z
datacite.date.available.fl_str_mv 2023-08-16T13:59:08Z
datacite.date.embargoed.fl_str_mv 2023-08-16T13:59:08Z
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datacite.subjects.subject.fl_str_mv CYP1A1
DNA strand breakage
Phenanthrene
Benzo[a]pyrene
Benzo[b]fluoranthene
datacite.titles.title.fl_str_mv Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
dc.contributor.none.fl_str_mv Repositório Científico de Acesso Aberto da ULisboa
dc.creator.none.fl_str_mv Bramatti, Isabella
Matos, Beatriz
Figueiredo, Neusa
Pousão-Ferreira, Pedro
Branco, Vasco
Martins, Marta
dc.date.Accepted.fl_str_mv 2023-01-01T00:00:00Z
dc.date.available.fl_str_mv 2023-08-16T13:59:08Z
dc.date.embargoed.fl_str_mv 2023-08-16T13:59:08Z
dc.format.none.fl_str_mv application/pdf
dc.identifier.none.fl_str_mv http://hdl.handle.net/10451/58891
dc.language.none.fl_str_mv eng
dc.publisher.none.fl_str_mv Elsevier
dc.rights.cclincense.fl_str_mv http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.rights.none.fl_str_mv http://purl.org/coar/access_right/c_abf2
dc.subject.none.fl_str_mv CYP1A1
DNA strand breakage
Phenanthrene
Benzo[a]pyrene
Benzo[b]fluoranthene
dc.title.fl_str_mv Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
dc.type.none.fl_str_mv http://purl.org/coar/resource_type/c_6501
description Polycyclic Aromatic Hydrocarbons (PAHs) are persistent pollutants normally found in the environment as complex mixtures. Although several individual PAHs are classified as mutagenic and carcinogenic pollutants, the interaction effects between compounds in a mixture may trigger different toxicological mechanisms and, consequently, yield different effects to organisms which are not accounted for in risk assessment guidelines. Given the ubiquity of PAHs, understanding the mechanistic features of their mixtures is a pressing research need. Therefore, the present work aimed to disclose the interaction effects of three PAHs with different carcinogenic potential and chemical structure, in primary hepatocyte cells of gilt-headed seabreams (Sparus aurata). Hepatocytes were exposed to Phenanthrene (Phe), Benzo[a]pyrene (B[a]P) and Benzo[b]fluoranthene (B[b]F) and their mixtures at different proportions and several cellular responses were analyzed: cellular viability, CYP1A1 activity (EROD assay) and protein expression level (Western blot); transcript (mRNA) levels of CYP1A1, EPXH1 and GST-3 (qRT-PCR); genotoxic effects (DNA strand breakage) by the Comet assay. Results show that B[a]P induced CYP1A1 gene and protein expression increasing its activity and, therefore, increasing the production of metabolites that trigger genotoxic DNA damage (%). Most importantly, mixtures containing Phe and B[a]P increased even further CYP1A1 mRNA levels and DNA damage (up to 70 %) which suggests that, although Phe is considered a non-carcinogenic PAH, it potentiates CYP1A1 synthesis induced by B[a]P, increasing its genotoxicity. These findings indicate that the upregulation of CYP1A1 by carcinogenic PAHs will not weaken even when in mixtures with non-carcinogenic PAHs. On contrary, non-carcinogenic PAHs may potentiate the genotoxic effect of carcinogenic PAH and therefore mixture composition should be taken in account when assessing PAH toxicity. In fact, our results point to the need of redefining Environmental Risk Assessment protocols for mixtures of carcinogenic pollutants.
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person_str_mv Bramatti, Isabella
Bramatti, Isabella
https://www.ciencia-id.pt/D41D-0E82-B973
D41D-0E82-B973
Matos, Beatriz
Matos, Beatriz
https://www.ciencia-id.pt/9418-6989-B909
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Figueiredo, Neusa
Figueiredo, Neusa
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Pousão-Ferreira, Pedro
Pousão-Ferreira, Pedro
https://www.ciencia-id.pt/5A11-AB29-0849
5A11-AB29-0849
http://orcid.org/0000-0001-6746-764X
0000-0001-6746-764X
Branco, Vasco
Branco, Vasco
https://www.ciencia-id.pt/671C-816B-F077
671C-816B-F077
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Martins, Marta
Martins, Marta
https://www.ciencia-id.pt/7810-1C0F-20D4
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http://orcid.org/0000-0002-3818-6256
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reponame_str Repositório da Universidade de Lisboa
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spelling engElsevierpt_PTPolycyclic Aromatic Hydrocarbons (PAHs) are persistent pollutants normally found in the environment as complex mixtures. Although several individual PAHs are classified as mutagenic and carcinogenic pollutants, the interaction effects between compounds in a mixture may trigger different toxicological mechanisms and, consequently, yield different effects to organisms which are not accounted for in risk assessment guidelines. Given the ubiquity of PAHs, understanding the mechanistic features of their mixtures is a pressing research need. Therefore, the present work aimed to disclose the interaction effects of three PAHs with different carcinogenic potential and chemical structure, in primary hepatocyte cells of gilt-headed seabreams (Sparus aurata). Hepatocytes were exposed to Phenanthrene (Phe), Benzo[a]pyrene (B[a]P) and Benzo[b]fluoranthene (B[b]F) and their mixtures at different proportions and several cellular responses were analyzed: cellular viability, CYP1A1 activity (EROD assay) and protein expression level (Western blot); transcript (mRNA) levels of CYP1A1, EPXH1 and GST-3 (qRT-PCR); genotoxic effects (DNA strand breakage) by the Comet assay. Results show that B[a]P induced CYP1A1 gene and protein expression increasing its activity and, therefore, increasing the production of metabolites that trigger genotoxic DNA damage (%). Most importantly, mixtures containing Phe and B[a]P increased even further CYP1A1 mRNA levels and DNA damage (up to 70 %) which suggests that, although Phe is considered a non-carcinogenic PAH, it potentiates CYP1A1 synthesis induced by B[a]P, increasing its genotoxicity. These findings indicate that the upregulation of CYP1A1 by carcinogenic PAHs will not weaken even when in mixtures with non-carcinogenic PAHs. On contrary, non-carcinogenic PAHs may potentiate the genotoxic effect of carcinogenic PAH and therefore mixture composition should be taken in account when assessing PAH toxicity. In fact, our results point to the need of redefining Environmental Risk Assessment protocols for mixtures of carcinogenic pollutants.application/pdfpt_PTInteraction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effectsPersonalBramatti, IsabellaDSpacehttp://dspace.org/items/5dcedc64-f9dd-4be1-b228-b5db6fc5cb95DSpacehttp://dspace.org/items/5dcedc64-f9dd-4be1-b228-b5db6fc5cb95Calvo BramattiIsabellaCiência IDhttps://www.ciencia-id.ptD41D-0E82-B973PersonalMatos, BeatrizDSpacehttp://dspace.org/items/9153b0eb-d3cd-4312-9c34-998f2a5c315bDSpacehttp://dspace.org/items/9153b0eb-d3cd-4312-9c34-998f2a5c315bManso de MatosBeatrizCiência IDhttps://www.ciencia-id.pt9418-6989-B909ORCIDhttp://orcid.org0000-0001-5923-5849PersonalFigueiredo, NeusaDSpacehttp://dspace.org/items/d2740563-4af4-406c-8e14-af87e66aca63DSpacehttp://dspace.org/items/d2740563-4af4-406c-8e14-af87e66aca63Loureiro Figueiredo MotubaneNeusa LindorosaCiência IDhttps://www.ciencia-id.pt341E-8FD2-22E2ORCIDhttp://orcid.org0000-0003-0062-1117PersonalPousão-Ferreira, PedroDSpacehttp://dspace.org/items/a239f428-e26c-4cc4-b490-f6caea01767eDSpacehttp://dspace.org/items/a239f428-e26c-4cc4-b490-f6caea01767eMarques Pousão-FerreiraPedroCiência IDhttps://www.ciencia-id.pt5A11-AB29-0849ORCIDhttp://orcid.org0000-0001-6746-764XScopus Author IDhttps://www.scopus.com6506349877PersonalBranco, VascoDSpacehttp://dspace.org/items/06d8e0eb-35e5-4d30-90d8-5af2fdab414bDSpacehttp://dspace.org/items/06d8e0eb-35e5-4d30-90d8-5af2fdab414bVeloso Neves BrancoVasco RuiCiência IDhttps://www.ciencia-id.pt671C-816B-F077ORCIDhttp://orcid.org0000-0002-0466-4934Scopus Author IDhttps://www.scopus.com8271087300PersonalMartins, MartaDSpacehttp://dspace.org/items/1edab515-0acc-4dc8-b94e-eb44cad2538cDSpacehttp://dspace.org/items/1edab515-0acc-4dc8-b94e-eb44cad2538cMartinsMartaCiência IDhttps://www.ciencia-id.pt7810-1C0F-20D4ORCIDhttp://orcid.org0000-0002-3818-6256Scopus Author IDhttps://www.scopus.com35603849900HostingInstitutionOrganizationalRepositório Científico de Acesso Aberto da ULisboae-mailmailto:repositorio@reitoria.ulisboa.ptrepositorio@reitoria.ulisboa.ptDOIIsPartOf10.1016/J.SCITOTENV.2022.1587832023-08-16T13:59:08Z20232022-11-22T12:08:52Z2023-01-01T00:00:00ZHandlehttp://hdl.handle.net/10451/58891http://purl.org/coar/access_right/c_abf2open accessCYP1A1DNA strand breakagePhenanthreneBenzo[a]pyreneBenzo[b]fluoranthene1364945 bytesFundação para a Ciência e a TecnologiaMixtures of Environmental Carcinogens: a molecular approach to improve environmental risk assessment strategies.3599-PPCDTCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaMarine and Environmental Sciences Centre6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaResearch Institute for Medicines6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaResearch Institute for Medicines6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaMolecular Insight on the Toxicity of Mixtures of Polycyclic Aromatic Hydrocarbons to Improve Environmental GuidelinesOECrossref Funder IDhttp://doi.org/10.13039/501100001871literaturehttp://purl.org/coar/resource_type/c_6501journal article2023http://creativecommons.org/licenses/by-nc-nd/4.0/http://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://repositorio.ulisboa.pt/bitstreams/5c9e585e-8ec1-4553-bf06-09307e832840/downloadScience of the Total Environment855158783
spellingShingle Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
Bramatti, Isabella
CYP1A1
DNA strand breakage
Phenanthrene
Benzo[a]pyrene
Benzo[b]fluoranthene
status SINGLETON
subject.fl_str_mv CYP1A1
DNA strand breakage
Phenanthrene
Benzo[a]pyrene
Benzo[b]fluoranthene
title Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
title_full Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
title_fullStr Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
title_full_unstemmed Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
title_short Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
title_sort Interaction of Polycyclic Aromatic Hydrocarbon compounds in fish primary hepatocytes: From molecular mechanisms to genotoxic effects
topic CYP1A1
DNA strand breakage
Phenanthrene
Benzo[a]pyrene
Benzo[b]fluoranthene
topic_facet CYP1A1
DNA strand breakage
Phenanthrene
Benzo[a]pyrene
Benzo[b]fluoranthene
url http://hdl.handle.net/10451/58891
visible 1