Mixtures derived from urban dust and diesel exhaust particles impact both estrogen-like signaling and estrogen-dependent cell proliferation in human breast cancer cell model

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F24%3AN0000189" target="_blank" >RIV/00027162:_____/24:N0000189 - isvavai.cz</a>

Výsledek na webu

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DOI - Digital Object Identifier

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Jazyk výsledku

angličtina

Název v původním jazyce

Mixtures derived from urban dust and diesel exhaust particles impact both estrogen-like signaling and estrogen-dependent cell proliferation in human breast cancer cell model

Popis výsledku v původním jazyce

Přednáška. Abstrakt v publikaci: XXXI. XENOBIOCHEMICKÉ SYMPOSIUM, 18. – 20. 9. 2024, Kyjov, ČR; ISBN 978-80-7672-050-3, [L13]. Airborne particulate matter (PM) represents a complex mixture of particles derived from combustion processes common in industry, traffic or local burning and heating. Chronic exposure to PM has been associated with diverse negative human health outcomes. An important group of pollutants associated with PM are polycyclic aromatic hydrocarbons (PAHs), as well as various of their derivatives, including polar compounds that are still poorly characterized. The general toxic modes of action of PAH include their well-known AhR-dependent genotoxicity, linked with production of mutagenic metabolites and formation of covalent DNA adducts, as well as a range of less defined non-genotoxic effects based on the interference with other intracellular signaling pathways. Increasing attention is being paid to the disruptive impact of PAHs on the estrogen receptor pathway signaling (ER), which might be linked to the activation of the AhR and the AhR-dependent metabolism of various polyaromatic compounds. Using both wild type (WT) and AhR-deficient variant (AhR KO) of estrogen-sensitive human breast cancer MCF- 7 cells, we studied effects of organic extracts (CE) and polar fractions (F3) of these extracts derived from reference standards representing urban dust (SRM 1649b) and diesel exhaust particles (SRM1650b). First, we evaluated their toxicity and genotoxicity by measuring cell viability and production of DNA adducts. Then, we analyzed the dysregulation of ER-related cellular processes. Finally, we assessed the modulation of ER-regulated genes and proteins. We found that relatively high doses of CE and F3 from both types of particles significantly reduced cell viability in both WT and AhR KO cells. Both CE and F3 formed minor levels of DNA adducts in WT cells. However, only SRM1650b-derived CE and F3 were able to induce DNA adducts in AhR KO cells. Both SRM1649b and 1650b stimulated ER signaling-related outputs, such as cell cycle progression and accumulation of cells in S-phase, ER-dependent cell proliferation, as well as the activation (phosphorylation) or induction of cell cycle related proteins, pRb (Ser 807/811) and Cyclin A2 in WT and AhR KO cells. We then assessed the capability of tested mixtures to bind and activate the ER reporter gene. The tested mixtures engaged the ER promoter and increased the transcription of the luciferase reporter gene. Notably, the luciferase activity was higher in WT cells than in AhR KO cells. Finally, we assessed the induction of the ER-regulated genes, TFF1 and PGR. Here, the expression of ER target genes was more prominent in the case of SRM1650b-derived CE and F3. Overall, our results suggest that polar compounds present in organic extracts of both types of complex PM mixtures may stimulate ER-dependent signaling in human cells. Most of these effects do not require active AhR-dependent metabolism. Interestingly, despite strong AhR activity of F3 fractions observed in our previous studies, this is not sufficient to repress ER activation in MCF-7 cells. Our results suggest that more attention should be paid to identification of PM-associated polar compounds with potential ER- disruptive activities.

Název v anglickém jazyce

Mixtures derived from urban dust and diesel exhaust particles impact both estrogen-like signaling and estrogen-dependent cell proliferation in human breast cancer cell model

Popis výsledku anglicky

Přednáška. Abstrakt v publikaci: XXXI. XENOBIOCHEMICKÉ SYMPOSIUM, 18. – 20. 9. 2024, Kyjov, ČR; ISBN 978-80-7672-050-3, [L13]. Airborne particulate matter (PM) represents a complex mixture of particles derived from combustion processes common in industry, traffic or local burning and heating. Chronic exposure to PM has been associated with diverse negative human health outcomes. An important group of pollutants associated with PM are polycyclic aromatic hydrocarbons (PAHs), as well as various of their derivatives, including polar compounds that are still poorly characterized. The general toxic modes of action of PAH include their well-known AhR-dependent genotoxicity, linked with production of mutagenic metabolites and formation of covalent DNA adducts, as well as a range of less defined non-genotoxic effects based on the interference with other intracellular signaling pathways. Increasing attention is being paid to the disruptive impact of PAHs on the estrogen receptor pathway signaling (ER), which might be linked to the activation of the AhR and the AhR-dependent metabolism of various polyaromatic compounds. Using both wild type (WT) and AhR-deficient variant (AhR KO) of estrogen-sensitive human breast cancer MCF- 7 cells, we studied effects of organic extracts (CE) and polar fractions (F3) of these extracts derived from reference standards representing urban dust (SRM 1649b) and diesel exhaust particles (SRM1650b). First, we evaluated their toxicity and genotoxicity by measuring cell viability and production of DNA adducts. Then, we analyzed the dysregulation of ER-related cellular processes. Finally, we assessed the modulation of ER-regulated genes and proteins. We found that relatively high doses of CE and F3 from both types of particles significantly reduced cell viability in both WT and AhR KO cells. Both CE and F3 formed minor levels of DNA adducts in WT cells. However, only SRM1650b-derived CE and F3 were able to induce DNA adducts in AhR KO cells. Both SRM1649b and 1650b stimulated ER signaling-related outputs, such as cell cycle progression and accumulation of cells in S-phase, ER-dependent cell proliferation, as well as the activation (phosphorylation) or induction of cell cycle related proteins, pRb (Ser 807/811) and Cyclin A2 in WT and AhR KO cells. We then assessed the capability of tested mixtures to bind and activate the ER reporter gene. The tested mixtures engaged the ER promoter and increased the transcription of the luciferase reporter gene. Notably, the luciferase activity was higher in WT cells than in AhR KO cells. Finally, we assessed the induction of the ER-regulated genes, TFF1 and PGR. Here, the expression of ER target genes was more prominent in the case of SRM1650b-derived CE and F3. Overall, our results suggest that polar compounds present in organic extracts of both types of complex PM mixtures may stimulate ER-dependent signaling in human cells. Most of these effects do not require active AhR-dependent metabolism. Interestingly, despite strong AhR activity of F3 fractions observed in our previous studies, this is not sufficient to repress ER activation in MCF-7 cells. Our results suggest that more attention should be paid to identification of PM-associated polar compounds with potential ER- disruptive activities.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

30108 - Toxicology

Projekt

<a href="/cs/project/GA21-00533S" target="_blank" >GA21-00533S: Nekonvenční environmentální ligandy Ah receptoru a jejich komplexní účinky in vitro</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Rok uplatnění

2024

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů