title: Investigation of the hepatotoxicity of flutamide: pro-survival/apoptotic and necrotic switch in primary rat hepatocytes characterized by metabolic and transcriptomic profiles in microfluidic liver biochips.
abstract: We investigated the effects of the liver damage induced by flutamide in primary rat hepatocytes using liver microfluidic biochips. Flutamide is a non-steroidal anti-androgenic drug. Two flutamide concentrations, 10 μM and 100 μM, were used to expose the hepatocytes for 24h under perfusion. Thanks to the maintenance of hepatocyte differentiation phenotype and to the biotransformation performance in the microfluidic cultures, the metabolic ratio analysis of hydroxyflutamide, flutamide-gluthatione and hydroxyflutamide-gluthatione productions demonstrated saturation of the drug's biotransformation process and the maintenance of a high level of flutamide at 100 μM when compared to 10 μM. A microarray analysis comparing flutamide (10 or 100 μM) with controls revealed a common response for both concentrations illustrated by modulating the expression of the mRNA of genes associated with mitochondrial perturbation, of the proliferator-activated receptors (Ppar) signaling, lipid and fatty acid metabolism, antioxidant defense, and cell death pathways, consistently with in vitro and in vivo reports. Additionally to literature reports, our integration of the transcriptomic profiles demonstrated a specific dose dependent response. We found at 10 μM a typical pro-survival/apoptosis network activation (through IGF/PDGFD upstream route and via a downstream up regulation in CREB5, BCL2, IKBKG routes in the PI3K/signaling). We also found a down regulation of mRNA levels in sugar and amino acid metabolism pathways. At 100 μM a typical necrosis switch was observed associated with a down regulation of the tight junctions' pathway, a cellular aggregation and a reduction of the cell viability. Altogether our data demonstrated the potential and the sensitivity of our liver microfluidic cultures to evaluate xenobiotic toxicity by improving in vitro analysis and reproducing both in vitro and in vivo results. Finally, we proposed two integrated synthetic networks to describe the response of rat hepatocytes to both exposure concentrations of flutamide.
authors.affiliation: CNRS UMR 7338, Laboratoire de Biomécanique et Bio ingénierie, Université de Technologie de Compiègne, France.
authors.forename: Audrey
authors.initials: A
authors.lastname: Legendre
authors.affiliation: INSERM U1016 Plate-forme génomique institut Cochin, 22 rue Méchain, 75014 Paris, France.
authors.forename: Sébastien
authors.initials: S
authors.lastname: Jacques
authors.affiliation: INSERM U1016 Plate-forme génomique institut Cochin, 22 rue Méchain, 75014 Paris, France.
authors.forename: Florent
authors.initials: F
authors.lastname: Dumont
authors.affiliation: Profilomic, 31 rue d'Aguesseau, 92100 Boulogne-Billancourt, France.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Cotton
authors.affiliation: CNRS UMR 7338, Laboratoire de Biomécanique et Bio ingénierie, Université de Technologie de Compiègne, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Paullier
authors.affiliation: CNRS UMR 7338, Laboratoire de Biomécanique et Bio ingénierie, Université de Technologie de Compiègne, France.
authors.forename: Marie José
authors.initials: MJ
authors.lastname: Fleury
authors.affiliation: CNRS UMR 7338, Laboratoire de Biomécanique et Bio ingénierie, Université de Technologie de Compiègne, France. Electronic address: eric.leclerc@utc.fr.
authors.forename: Eric
authors.initials: E
authors.lastname: Leclerc
mesh_terms: D000726:Androgen Antagonists / Q000633:toxicity*; D000818:Animals; D019149:Bioreactors; D002478:Cells, Cultured; D019363:Cytochrome P-450 CYP1A1 / Q000378:metabolism; D005485:Flutamide / Q000633:toxicity*; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D046228:Microarray Analysis; D046210:Microfluidic Analytical Techniques; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats
chemical_list: D000726:Androgen Antagonists; D012333:RNA, Messenger; D005485:Flutamide; D019363:Cytochrome P-450 CYP1A1
keywords: Flutamide; Hepatotoxicity; Metabolism; Microarray; Microfluidic biochips; Rat primary hepatocytes; Transcriptomic