Functional expression, inhibition and induction of CYP enzymes in HepaRG cells

Turpeinen, Miia; Tolonen, Ari; Chesné, Christophe; Guillouzo, André; Uusitalo, Jouko; Pelkonen, Olavi

doi:10.1016/j.tiv.2009.03.008

Cited by 99 publications

(56 citation statements)

References 24 publications

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“…In particular, they are able to express numerous P450 cytochromes, allowing the performance of many normal metabolic liver functions such as the production of phase I and II enzymes and transmembrane transport proteins, unlike other hepatocyte cell lines such as HepG2 (Guillouzo et al 2007;Kanebratt and Andersson 2008;Turpeinen et al 2009;Jennen et al 2010). …”

Section: Discussionmentioning

confidence: 99%

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

et al. 2017

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Magnetic mesoporous silica nanoparticles (M-MSNs) are a promising class of nanoparticles for drug delivery. However, a deep understanding of the toxicological mechanisms of action of these nanocarriers is essential, especially in the liver. The potential toxicity on HepaRG cells of pristine, pegylated (PEG), and lipid (DMPC) M-MSNs were compared. Based on MTT assay and real-time cell impedance, none of these NPs presented an extensive toxicity on hepatic cells. However, we observed by transmission electron microscopy (TEM) that the DMPC and pristine M-MSNs were greatly internalized. In comparison, PEG M-MSNs showed a slower cellular uptake. Whole gene expression profiling revealed the M-MSNs molecular modes of action in a time-and dose-dependent manner. The lowest dose tested (1.6 mg/cm 2 ) induced no molecular effect and was defined as 'No Observed Transcriptional Effect level.' The dose 16 mg/cm 2 revealed nascent but transient effects. At the highest dose (80 mg/cm 2 ), adverse effects have clearly arisen and increased over time. The limit of biocompatibility for HepaRG cells could be set at 16 mg/cm 2 for these NPs. Thanks to a comparative pathway-driven analysis, we highlighted the sequence of events that leads to the disruption of hepatobiliary system, elicited by the three types of MMSNs, at the highest dose. The Adverse Outcome Pathway of hepatic cholestasis was implicated. Toxicogenomics applied to cell cultures is an effective tool to characterize and compare the modes of action of many substances. We propose this strategy as an asset for upstream selection of the safest nanocarriers in the framework of regulation for nanobiosafety. ARTICLE HISTORY

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Section: Discussionmentioning

confidence: 99%

Biocompatibility assessment of functionalized magnetic mesoporous silica nanoparticles in human HepaRG cells

et al. 2017

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“…Differentiated HepaRG cells express various liver functions, including P450s, phase II enzymes, transporters, and nuclear receptors at levels comparable with those found in primary hepatocytes and are responsive to prototypical inducers, suggesting that they could represent a surrogate to the latter in drug metabolism and toxicity studies (Aninat et al, 2006;Le Vee et al, 2006;Guillouzo et al, 2007;Josse et al, 2008;Kanebratt and Andersson, 2008b;Turpeinen et al, 2009). Moreover, some evidence has been provided that LD-seeded HepaRG cells can retain relatively stable expression and activities of P450s for several weeks at confluence (Josse et al, 2008;Kanebratt and Andersson, 2008a).…”

mentioning

confidence: 99%

Stable Expression, Activity, and Inducibility of Cytochromes P450 in Differentiated HepaRG Cells

Anthérieu

Chesné²,

Li³

et al. 2009

Drug Metab Dispos

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230

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ABSTRACT:HepaRG cells possess the unique property to differentiate in vitro and to express various functions of mature hepatocytes, including the major cytochromes P450 (P450s). In the present study, we carefully analyzed mRNA expression and activity of the major P450s and their responsiveness to three prototypical inducers, phenobarbital, rifampicin, and omeprazole, in differentiated HepaRG cell cultures over a 4-week period after low and high seeding. Only minor differences were observed in P450 activities when measured by two cocktails of probe substrates, probably related to the choice and/or concentration of substrates. Similar results were obtained from the two cell seeding conditions. Expression and activities of several P450s were dimethyl sulfoxidedependent. However, basal P450 expression and activities as well as their responsiveness to the prototypical inducers were well maintained over the 4-week period, and a good correlation was observed between transcript levels and corresponding activities. Thus, CYP1A2, CYP2B6, and CYP3A4 were found to accurately respond to their respective prototypical inducers, i.e., omeprazole, phenobarbital, and rifampicin. Likewise, basal expression of several phase II enzymes, transporters, and nuclear receptors, and response to inducers were also well preserved. More genes were found to be induced in HepaRG cells than in primary human hepatocytes, and no marked variation was noticed between the different passages. Taken together, these data support the conclusion that HepaRG cells represent a promising surrogate to primary human hepatocytes for xenobiotic metabolism and toxicity studies.

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“…Likewise, HepaRG cells express a large panel of liver-specific genes including several CYP enzymes, which is in contrast to HepG2 cell lines. In addition to P450 enzymes, HepaRG cells have a stable expression of phase II enzymes, transporters and nuclear transcription factors over a time period of six weeks in culture (Aninat et al 2006;Anthérieu et al 2010;Kanebratt and Andersson, 2008;Turpeinen et al 2009). Both immunoblotting and reverse transcription polymerase chain reaction (RT-PCR) techniques have been applied to examine the pesticide-CYP induction (Wyde et al 2003;Lemaire et al 2004;Das et al 2006;Sun et al 2005;Johri et al 2007;Barber et al 2007).…”

Section: Induction Of Cyp Enzymesmentioning

confidence: 99%