Cellular stress responses as novel in vitro toxicity markers of selected polycyclic aromatic hydrocarbons

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F20%3AN0000212" target="_blank" >RIV/00027162:_____/20:N0000212 - 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

Cellular stress responses as novel in vitro toxicity markers of selected polycyclic aromatic hydrocarbons

Popis výsledku v původním jazyce

We established a novel complex in vitro system for testing of acute toxicity responses induced by various xenobiotics. The system is based on qRT-PCR analysis of expression profiles of genes that are involved in cellular stress responses, including: early stress response; heat shock protein (HSP) response; DNA damage response; unfolded protein response (UPR; related to endoplasmic reticulum (ER) stress); and immunotoxicity. The cellular stress markers represent integral parameters affected by various intracellular pathways and they are complementary to e.g. luciferase reporter gene (CALUX) assays detecting specific nuclear receptor activation. Ubiquitous environmental pollutants, polycyclic aromatic hydrocarbons (PAHs) represent a diverse group of organic substances exerting different types of toxicities, such as activation of the aryl hydrocarbon receptor (AhR) and/or genotoxicity. In this study, we screened cellular stress markers, in order to detect non-conventional modes of action of PAHs, and compare them with their other well-recognized toxic effects. Six individual PAHs with unique characteristics were selected for the study: fluoranthene (Fla), pyrene (Pyr), chrysene (Chry), benzo[a]anthracene (BaA), benzo[a]pyrene (BaP), and benzo[k]fluoranthene (BkF). Differentiated human liver HepaRG cells were exposed to the selected PAHs for 24 h, and changes of mRNA levels of genes involved in early stress (EGR1, ATF3, GDF15), DNA damage (CDKN1A), HSP response (HSP70), ER stress (HSPA, DDIT3) and immunotoxicity (interleukin 8) were detected. The results were analysed with respect to the effects of model substances, e.g. thapsigargin, ER-stress inducer, or lipopolysaccharide. We found that CDKN1A and early stress genes ATF3 and EGR1 were induced by BaP and BkF. Moreover, GDF15 mRNA was increased by Chry, BaA, BaP and BkF. Expression of other cellular stress markers was not affected by carcinogenic PAHs. Low-molecular-weight PAHs, Fla and Pyr, did not induce cellular stress markers. Determination of stress markers in the cells exposed to xenobiotics such as environmental contaminants or their mixtures is a novel approach suitable for in vitro toxicity profiling. The system represents a sensitive first-line set of parameters suitable for identification of in vitro toxicity, complementing the general cytotoxicity assays.

Název v anglickém jazyce

Cellular stress responses as novel in vitro toxicity markers of selected polycyclic aromatic hydrocarbons

Popis výsledku anglicky

We established a novel complex in vitro system for testing of acute toxicity responses induced by various xenobiotics. The system is based on qRT-PCR analysis of expression profiles of genes that are involved in cellular stress responses, including: early stress response; heat shock protein (HSP) response; DNA damage response; unfolded protein response (UPR; related to endoplasmic reticulum (ER) stress); and immunotoxicity. The cellular stress markers represent integral parameters affected by various intracellular pathways and they are complementary to e.g. luciferase reporter gene (CALUX) assays detecting specific nuclear receptor activation. Ubiquitous environmental pollutants, polycyclic aromatic hydrocarbons (PAHs) represent a diverse group of organic substances exerting different types of toxicities, such as activation of the aryl hydrocarbon receptor (AhR) and/or genotoxicity. In this study, we screened cellular stress markers, in order to detect non-conventional modes of action of PAHs, and compare them with their other well-recognized toxic effects. Six individual PAHs with unique characteristics were selected for the study: fluoranthene (Fla), pyrene (Pyr), chrysene (Chry), benzo[a]anthracene (BaA), benzo[a]pyrene (BaP), and benzo[k]fluoranthene (BkF). Differentiated human liver HepaRG cells were exposed to the selected PAHs for 24 h, and changes of mRNA levels of genes involved in early stress (EGR1, ATF3, GDF15), DNA damage (CDKN1A), HSP response (HSP70), ER stress (HSPA, DDIT3) and immunotoxicity (interleukin 8) were detected. The results were analysed with respect to the effects of model substances, e.g. thapsigargin, ER-stress inducer, or lipopolysaccharide. We found that CDKN1A and early stress genes ATF3 and EGR1 were induced by BaP and BkF. Moreover, GDF15 mRNA was increased by Chry, BaA, BaP and BkF. Expression of other cellular stress markers was not affected by carcinogenic PAHs. Low-molecular-weight PAHs, Fla and Pyr, did not induce cellular stress markers. Determination of stress markers in the cells exposed to xenobiotics such as environmental contaminants or their mixtures is a novel approach suitable for in vitro toxicity profiling. The system represents a sensitive first-line set of parameters suitable for identification of in vitro toxicity, complementing the general cytotoxicity assays.

Druh

O - Ostatní výsledky

CEP obor

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OECD FORD obor

30108 - Toxicology

Projekt

<a href="/cs/project/EF15_003%2F0000495" target="_blank" >EF15_003/0000495: FIT (Farmakologie, Imunoterapie, nanoToxikologie)</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2020

Kód důvěrnosti údajů

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