title: Induction of expression of aryl hydrocarbon receptor-dependent genes in human HepaRG cell line modified by shRNA and treated with β-naphthoflavone.
abstract: The aryl hydrocarbon receptor (AhR) mediates a variety of biological responses to ubiquitous environmental pollutants. In this study, the effects of administration of β-naphthoflavone (BNF), a potent AhR ligand, on the expression of AhR-dependent genes were examined by microarray and qPCR analysis in both, differentiated and undifferentiated HepaRG cell lines. To prove that BNF-induced changes of investigated genes were indeed AhR-dependent, we knock down the expression of AhR by stable transfection of HepaRG cells with shRNA. Regardless of genetical identity, our results clearly demonstrate different expression profiles of AhR-dependent genes between differentiated and undifferentiated HepaRG cells. Genes involved in metabolism of xenobiotics constitute only minute fraction of all genes regulated by AhR in HepaRG cells. Participation of AhR in induction of expression of genes associated with regulation of apoptosis or involved in cell proliferation as well as AhR-dependent inhibition of genes connected to cell adhesion could support suggestion of involvement of AhR not only in initiation but also in progression of carcinogenesis. Among the AhR-dependent genes known to be involved in metabolism of xenobiotics, cytochromes P4501A1 and 1B1 belong to the most inducible by BNF. On the contrary, expression of GSTA1 and GSTA2 was significantly inhibited after BNF treatment of HepaRG cells. Among the AhR-dependent genes that are not involved in metabolism of xenobiotics SERPINB2, STC2, ARL4C, and TIPARP belong to the most inducible by BNF. Our results imply involvement of Ah receptor in regulation of CYP19A1, the gene-encoding aromatase, and an enzyme responsible for a key step in the biosynthesis of estrogens.
authors: Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479, Poznan, Poland. brauzdam@man.poznan.pl.
authors: Damian
authors: http://orcid.org/0000-0001-8868-4897
authors: D
authors: Brauze
authors: Department of Histology and Embryology, Poznan University of Medical Sciences, 60-781, Poznan, Poland.|Department of Anatomy, Poznań University of Medical Sciences, 60-781, Poznan, Poland.
authors: Piotr
authors: P
authors: Zawierucha
authors: Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479, Poznan, Poland.
authors: Katarzyna
authors: K
authors: Kiwerska
authors: Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479, Poznan, Poland.
authors: Kinga
authors: K
authors: Bednarek
authors: Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479, Poznan, Poland.
authors: Martyna
authors: M
authors: Oleszak
authors: Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479, Poznan, Poland.|Department of Medical Genetics, Medical University of Warsaw, Pawinskiego 3c, 02-106, Warsaw, Poland.
authors: Malgorzata
authors: M
authors: Rydzanicz
authors: Institute of Human Genetics, Polish Academy of Sciences, Strzeszynska 32, 60-479, Poznan, Poland.
authors: Malgorzata
authors: M
authors: Jarmuz-Szymczak
mesh_terms: D051792:Basic Helix-Loop-Helix Proteins / Q000096:biosynthesis* / Q000235:genetics; D002448:Cell Adhesion / Q000187:drug effects; D002460:Cell Line; D004967:Estrogens / Q000096:biosynthesis / Q000235:genetics; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D034741:RNA, Small Interfering / Q000235:genetics / Q000378:metabolism*; D018336:Receptors, Aryl Hydrocarbon / Q000096:biosynthesis* / Q000235:genetics; D019324:beta-Naphthoflavone / Q000494:pharmacology*
chemical_list: D051792:Basic Helix-Loop-Helix Proteins; D004967:Estrogens; D034741:RNA, Small Interfering; D018336:Receptors, Aryl Hydrocarbon; D019324:beta-Naphthoflavone; C111235:AHR protein, human
keywords: ARL4C; AhR; CCNE2; CYP19A1 SERPINB2; CYP1A1; CYP1A2; CYP1B1; GSTA2; HepaRG cell line; NQO1; SLC7A5; STC2; TIPARP; β-Naphthoflavone