Genomic and phenotype changes of human bronchial epithelial cells during benzo[a]pyrene-induced transformation

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F22%3AN0000078" target="_blank" >RIV/00027162:_____/22:N0000078 - 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

Genomic and phenotype changes of human bronchial epithelial cells during benzo[a]pyrene-induced transformation

Popis výsledku v původním jazyce

Polycyclic aromatic hydrocarbons (PAHs) and their derivatives belong among principle air pollutants, which elicit both genotoxic and non-genotoxic effects. One of the most studied carcinogenic PAHs is benzo[a]pyrene (BaP), a potent agonist of the aryl hydrocarbon receptor (AhR), which plays an important role in many processes associated with cancer promotion and/or progression. In this study, we used normal human bronchial epithelial HBEC-12KT cells and their derivative - BaP-transformed HBEC-12KT-B1 cells (cells were cultivated in presence of 1 µM BaP for 12 weeks). These cells underwent epithelial-to-mesenchymal transition (EMT), gained mesenchymal-like phenotype, started to migrate and adopt expression features characteristic for EMT such as decreased expression of E-cadherin or increased expression of N-cadherin, vimentin, TWIST or ZEB1. We investigated the kinetics of transcriptional and phenotypical changes during BaP-induced cellular transformation of the HBEC-12KT cells in both short (24-72 hours) and prolonged (2-10 weeks) exposure times. First, we performed a series of microarray analyses in order to delineate the transcriptional background of early morphological changes induced by BaP after 2 weeks and compared them with those accompanying acute exposure of HBEC-12KT cells to BaP for 24 h. Due to the fact that BaP is a potent agonist of the AhR, next part of this study was dedicated to answer if another AhR agonist, herein we chose 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), may also induce cellular transformation of the HBEC-12KT model. Interestingly, TCDD did not induce cellular transformation even after 10 weeks of repeated exposure. Transcriptional changes were compared between cells exposed to BaP and cells exposed to TCDD for 24 h or 2 weeks, and led us to identification of gene set, whose expression is regulated by AhR as it was significantly compromised in cells exposed to TCDD and CH223191, an AhR inhibitor. However, AhR activation was only a prerequisite of EMT and its activation alone was not sufficient to reduce EMT process. Final part of our study was focused on identification of transcriptional and functional features preceding establishment of EMT-like phenotype. Upregulation of p21, Notch signaling and CXCL5 production and secretion were found as deregulated in both HBEC-12KT cells exposed to BaP for 2 weeks and remained so in terminally transformed HBEC-12KT-B1 cells.

Název v anglickém jazyce

Genomic and phenotype changes of human bronchial epithelial cells during benzo[a]pyrene-induced transformation

Popis výsledku anglicky

Polycyclic aromatic hydrocarbons (PAHs) and their derivatives belong among principle air pollutants, which elicit both genotoxic and non-genotoxic effects. One of the most studied carcinogenic PAHs is benzo[a]pyrene (BaP), a potent agonist of the aryl hydrocarbon receptor (AhR), which plays an important role in many processes associated with cancer promotion and/or progression. In this study, we used normal human bronchial epithelial HBEC-12KT cells and their derivative - BaP-transformed HBEC-12KT-B1 cells (cells were cultivated in presence of 1 µM BaP for 12 weeks). These cells underwent epithelial-to-mesenchymal transition (EMT), gained mesenchymal-like phenotype, started to migrate and adopt expression features characteristic for EMT such as decreased expression of E-cadherin or increased expression of N-cadherin, vimentin, TWIST or ZEB1. We investigated the kinetics of transcriptional and phenotypical changes during BaP-induced cellular transformation of the HBEC-12KT cells in both short (24-72 hours) and prolonged (2-10 weeks) exposure times. First, we performed a series of microarray analyses in order to delineate the transcriptional background of early morphological changes induced by BaP after 2 weeks and compared them with those accompanying acute exposure of HBEC-12KT cells to BaP for 24 h. Due to the fact that BaP is a potent agonist of the AhR, next part of this study was dedicated to answer if another AhR agonist, herein we chose 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), may also induce cellular transformation of the HBEC-12KT model. Interestingly, TCDD did not induce cellular transformation even after 10 weeks of repeated exposure. Transcriptional changes were compared between cells exposed to BaP and cells exposed to TCDD for 24 h or 2 weeks, and led us to identification of gene set, whose expression is regulated by AhR as it was significantly compromised in cells exposed to TCDD and CH223191, an AhR inhibitor. However, AhR activation was only a prerequisite of EMT and its activation alone was not sufficient to reduce EMT process. Final part of our study was focused on identification of transcriptional and functional features preceding establishment of EMT-like phenotype. Upregulation of p21, Notch signaling and CXCL5 production and secretion were found as deregulated in both HBEC-12KT cells exposed to BaP for 2 weeks and remained so in terminally transformed HBEC-12KT-B1 cells.

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

2022

Kód důvěrnosti údajů

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