A simultaneous assessment of CYP3A4 metabolism and induction in the DPX-2 cell line

Trubetskoy, Olga V.; Marks, Bryan D.; Zielinski, Thomas; Yueh, Mei‐Fei; Raucy, Judy L.

doi:10.1208/aapsj070102

Cited by 48 publications

(35 citation statements)

References 33 publications

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“…2 and 3). Among the compounds tested, rifampicin, bosentan, moricizine, and ritonavir are reported as selective PXR-activators (LeCluyse, 2001;Luo et al, 2002;van Giersbergen et al, 2002), phenobarbital, phenytoin, and efavirenz are PXR/CAR dual activators (Hariparsad et al, 2004;Wang et al, 2004;Trubetskoy et al, 2005;Bell and Michalopoulos, 2006;Faucette et al, 2007), and CITCO and artemisinin are selective CAR activators (Maglich et al, 2003;Simonsson et al, 2006). Rifampicin-mediated CYP3A4 induction was conducted in Fa2N-4 cells and four batches of human hepatocytes (DMQ, 527, 455, and LHO).…”

Section: Resultsmentioning

confidence: 99%

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Comparison of Immortalized Fa2N-4 Cells and Human Hepatocytes as in Vitro Models for Cytochrome P450 Induction

Hariparsad¹,

Carr²,

Evers³

et al. 2008

Drug Metab Dispos

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ABSTRACT:Fa2N-4 cells have been proposed as a tool to identify CYP3A4 inducers. To evaluate whether Fa2N-4 cells are a reliable surrogate for cryopreserved human hepatocytes, we assessed the basal mRNA expression of 64 drug disposition genes in Fa2N-4 cells. Significant differences were found in the expression of major drugmetabolizing enzymes, nuclear receptors, and transporters between both cell types. Importantly, the expression of constitutive androstane receptor (CAR) and several hepatic uptake transporters was significantly lower (>50-fold) in Fa2N-4 cells, whereas the expression of pregnane X-receptor (PXR) and aryl hydrocarbon receptor (

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Section: Resultsmentioning

confidence: 99%

“…b Gene induction occurs via cross-talk between PXR and CAR (Wang et al, 2004;Trubetskoy et al, 2005;Bell and Michalopoulos, 2006). c Dose-response curve did not reach saturation for CYP3A4 induction within concentrations tested.…”

Section: Discussionmentioning

confidence: 99%

Comparison of Immortalized Fa2N-4 Cells and Human Hepatocytes as in Vitro Models for Cytochrome P450 Induction

Hariparsad¹,

Carr²,

Evers³

et al. 2008

Drug Metab Dispos

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“…Currently, the cell line most often used in CYP3A4 reporter gene assays is the human hepatocarcinoma-derived HepG2 cell line (Ogg et al, 1997(Ogg et al, , 1999El-Sankary et al, 2001;Luo et al, 2004;Trubetskoy et al, 2005;Noracharttiyapot et al, 2006;Sinz et al, 2006;Huang et al, 2007). However, HepG2 mainly expresses the fetal enzyme CYP3A7 instead of CYP3A4 (Schuetz et al, 1993).…”

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confidence: 99%

Comparison of Two Immortalized Human Cell Lines to Study Nuclear Receptor-Mediated CYP3A4 Induction

Harmsen¹,

Koster²,

Beijnen³

et al. 2008

Drug Metab Dispos

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ABSTRACT:Since CYP3A4 is responsible for the biotransformation of over 50% of all clinically used drugs, induction results in an increased clearance of many concomitantly administered drugs, thereby decreasing treatment efficacy or, in the case of prodrugs, lead to severe intoxications. CYP3A4 induction is regulated by the pregnane X receptor, constitutive androstane receptor, and vitamin D receptor. Since these nuclear receptors show large interspecies differences, accurate prediction of nuclear receptor-mediated CYP3A4 induction in humans requires the use of human systems. Because primary cultures of human hepatocytes or enterocytes have major drawbacks like poor availability and poor reproducibility, human cell lines are a good alternative. In this study, the widely used HepG2 cell line was compared with the LS180 cell line to serve as a model to study CYP3A4 induction. There was a clear difference between the cell lines with respect to CYP3A enzyme expression and induction. In LS180, CYP3A4 was expressed and was found to be induced by prototypical nuclear receptor agonists, whereas in HepG2, CYP3A4 was nonresponsive to treatment with rifampicin, CITCO [6-(4-chlorophenyl)imidazo[2,1-b][1,3]thiazole-5-carbaldehyde-O-3,4-dichlorobenzyl) oxime], or calcitriol. We subsequently evaluated whether these host-cell differences also have an effect on CYP3A4 reporter gene activity. We clearly show that there are differences in CYP3A4 reporter activity between the cell lines, and based on these results and those found on mRNA and protein level, we conclude that LS180 is a more suitable cell line to study CYP3A4 induction than the widely used HepG2.

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“…All water solutions, which contained 10% w/v rubusoside, were diluted 100-fold in water to a final rubusoside concentration of 0.1%. In previous studies, DMso concentrations as low as 0.1% have been used to solubilize paclitaxel (20). therefore, a parallel set of ptX solutions (1-10 µg/ml) in 2 µg/ml cpt solubilized by 100% DMso were prepared and then were each diluted 100-fold with water to a final DMSO concentration of 1% v/v.…”

Section: Methodsmentioning

confidence: 99%

Paclitaxel-induced apoptosis is blocked by camptothecin in human breast and pancreatic cancer cells

Jeansonne

Koh²,

Zhang³

et al. 2011

Oncol Rep

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Abstract. the combination of paclitaxel (ptX) and topoisomerase I inhibitors such as camptothecin (cpt) constitutes a therapeutic strategy based on anticipated synergism. However, previous in vitro studies have generated contradictory findings for this strategy. The interaction between these drugs can be synergistic or antagonistic, depending on the cell type examined. to gain additional insight into this promising yet controversial strategy, we investigated the interaction between ptX and cpt in three different cell lines (pAnc-1, MDA-MB-231 and Hl-60) and explored possible underlying mechanisms of synergy or antagonism. Using a novel solubilizing natural compound, rubusoside, water-insoluble ptX and cpt were solubilized to enable the comparison of the effects of single drugs and their combination on cell viability. Intracellular drug concentrations were quantified to examine the effect of CPT on cellular uptake and accumulation of ptX. Flow cytometry and quantitative real-time pcr gene array analyses were used to explore the mechanisms behind the interaction between ptX and cpt. Our studies confirmed that rubusoside-solubilized PTX or CPT maintained cytotoxicity, causing significant reductions in cell viability. However, the efficacy of the combination of ptX and cpt produced varied results based on the cell line tested. cpt antagonistically reduced the cytotoxic activity of ptX in pAnc-1 and MDA-MB-231 cells. the effect of cpt on the cytotoxicity of ptX was less pronounced in Hl-60 cells, showing neither synergy nor antagonism. Analysis of apoptosis by flow cytometry revealed that upon co-treatment with cpt, apoptosis induced by ptX was attenuated in pAnc-1 and MDA-MB-231 cells. In agreement with our cytotoxicity findings, no synergistic or antagonistic effects on apoptosis were observed in Hl-60 cells. the antagonism in pAnc-1 and MDA-MB-231 cells was not a result of reduced PTX uptake and accumulation because the amount of intracellular ptX was not altered upon co-treatment with cpt. Moreover, higher expression of anti-apoptosisrelated transcripts (BCL2L10, CFLAR, HIP1 and TRADD) in pAnc-1 cells was observed upon combination treatment over ptX treatment alone. Although exact underlying mechanisms are unknown, the suspected CPT-dependent reduction of intracellular PTX accumulation was ruled out. The findings of antagonism and increased anti-apoptotic gene transcription serve as a precaution to the design of combination drug strategies where a synergistic interaction may not exist.

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A simultaneous assessment of CYP3A4 metabolism and induction in the DPX-2 cell line

Cited by 48 publications

References 33 publications

Comparison of Immortalized Fa2N-4 Cells and Human Hepatocytes as in Vitro Models for Cytochrome P450 Induction

Comparison of Immortalized Fa2N-4 Cells and Human Hepatocytes as in Vitro Models for Cytochrome P450 Induction

Comparison of Two Immortalized Human Cell Lines to Study Nuclear Receptor-Mediated CYP3A4 Induction

Paclitaxel-induced apoptosis is blocked by camptothecin in human breast and pancreatic cancer cells

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