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Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity

Human hepatoma HepaRG cells express most drug metabolizing enzymes and constitute a pertinent in vitro alternative cell system to primary cultures of human hepatocytes in order to determine drug metabolism and evaluate the toxicity of xenobiotics. In this work, we established novel transgenic HepaRG...

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Autores principales: Vlach, Manuel, Quesnot, Nicolas, Dubois-Pot-Schneider, Hélène, Ribault, Catherine, Verres, Yann, Petitjean, Kilian, Rauch, Claudine, Morel, Fabrice, Robin, Marie-Anne, Corlu, Anne, Loyer, Pascal
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6567340/
https://www.ncbi.nlm.nih.gov/pubmed/31096615
http://dx.doi.org/10.3390/s19102245
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author Vlach, Manuel
Quesnot, Nicolas
Dubois-Pot-Schneider, Hélène
Ribault, Catherine
Verres, Yann
Petitjean, Kilian
Rauch, Claudine
Morel, Fabrice
Robin, Marie-Anne
Corlu, Anne
Loyer, Pascal
author_facet Vlach, Manuel
Quesnot, Nicolas
Dubois-Pot-Schneider, Hélène
Ribault, Catherine
Verres, Yann
Petitjean, Kilian
Rauch, Claudine
Morel, Fabrice
Robin, Marie-Anne
Corlu, Anne
Loyer, Pascal
author_sort Vlach, Manuel
collection PubMed
description Human hepatoma HepaRG cells express most drug metabolizing enzymes and constitute a pertinent in vitro alternative cell system to primary cultures of human hepatocytes in order to determine drug metabolism and evaluate the toxicity of xenobiotics. In this work, we established novel transgenic HepaRG cells transduced with lentiviruses encoding the reporter green fluorescent protein (GFP) transcriptionally regulated by promoter sequences of cytochromes P450 (CYP) 1A1/2, 2B6 and 3A4 genes. Here, we demonstrated that GFP-biosensor transgenes shared similar expression patterns with the corresponding endogenous CYP genes during proliferation and differentiation in HepaRG cells. Interestingly, differentiated hepatocyte-like HepaRG cells expressed GFP at higher levels than cholangiocyte-like cells. Despite weaker inductions of GFP expression compared to the strong increases in mRNA levels of endogenous genes, we also demonstrated that the biosensor transgenes were induced by prototypical drug inducers benzo(a)pyrene and phenobarbital. In addition, we used the differentiated biosensor HepaRG cells to evidence that pesticide mancozeb triggered selective cytotoxicity of hepatocyte-like cells. Our data demonstrate that these new biosensor HepaRG cells have potential applications in the field of chemicals safety evaluation and the assessment of drug hepatotoxicity.
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spelling pubmed-65673402019-06-17 Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity Vlach, Manuel Quesnot, Nicolas Dubois-Pot-Schneider, Hélène Ribault, Catherine Verres, Yann Petitjean, Kilian Rauch, Claudine Morel, Fabrice Robin, Marie-Anne Corlu, Anne Loyer, Pascal Sensors (Basel) Article Human hepatoma HepaRG cells express most drug metabolizing enzymes and constitute a pertinent in vitro alternative cell system to primary cultures of human hepatocytes in order to determine drug metabolism and evaluate the toxicity of xenobiotics. In this work, we established novel transgenic HepaRG cells transduced with lentiviruses encoding the reporter green fluorescent protein (GFP) transcriptionally regulated by promoter sequences of cytochromes P450 (CYP) 1A1/2, 2B6 and 3A4 genes. Here, we demonstrated that GFP-biosensor transgenes shared similar expression patterns with the corresponding endogenous CYP genes during proliferation and differentiation in HepaRG cells. Interestingly, differentiated hepatocyte-like HepaRG cells expressed GFP at higher levels than cholangiocyte-like cells. Despite weaker inductions of GFP expression compared to the strong increases in mRNA levels of endogenous genes, we also demonstrated that the biosensor transgenes were induced by prototypical drug inducers benzo(a)pyrene and phenobarbital. In addition, we used the differentiated biosensor HepaRG cells to evidence that pesticide mancozeb triggered selective cytotoxicity of hepatocyte-like cells. Our data demonstrate that these new biosensor HepaRG cells have potential applications in the field of chemicals safety evaluation and the assessment of drug hepatotoxicity. MDPI 2019-05-15 /pmc/articles/PMC6567340/ /pubmed/31096615 http://dx.doi.org/10.3390/s19102245 Text en © 2019 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Vlach, Manuel
Quesnot, Nicolas
Dubois-Pot-Schneider, Hélène
Ribault, Catherine
Verres, Yann
Petitjean, Kilian
Rauch, Claudine
Morel, Fabrice
Robin, Marie-Anne
Corlu, Anne
Loyer, Pascal
Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
title Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
title_full Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
title_fullStr Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
title_full_unstemmed Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
title_short Cytochrome P450 1A1/2, 2B6 and 3A4 HepaRG Cell-Based Biosensors to Monitor Hepatocyte Differentiation, Drug Metabolism and Toxicity
title_sort cytochrome p450 1a1/2, 2b6 and 3a4 heparg cell-based biosensors to monitor hepatocyte differentiation, drug metabolism and toxicity
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6567340/
https://www.ncbi.nlm.nih.gov/pubmed/31096615
http://dx.doi.org/10.3390/s19102245
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