Up-Regulation of Human CYP2J2 in HepG2 Cells by Butylated Hydroxyanisole Is Mediated by c-Jun and Nrf2

Lee, Andy Chao Hsuan; Murray, Michael T.

doi:10.1124/mol.109.062729

Cited by 26 publications

(20 citation statements)

References 34 publications

(52 reference statements)

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“…1A). The isothiocyanate, SFN, is believed to be a classic activator of only Nrf2 (Nguyen et al, 2009;Ahn et al, 2010), whereas BHA and acrolein modulate gene expression by activation of either Nrf2 or AP-1 (Choi and Moore, 1993;Lee and Murray, 2010). As anticipated, HO-1 (Ϸ5-fold) and NQO-1 (Ϸ3-fold) mRNA levels were significantly induced by SFN, but no significant induction of Aldh1a1 gene expression was observed with SFN ( Fig.…”

Section: Methodssupporting

confidence: 49%

See 1 more Smart Citation

Activator Protein-1 Regulation of Murine Aldehyde Dehydrogenase 1a1

Makia

Amunom²,

Falkner³

et al. 2012

Mol Pharmacol

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Previously we demonstrated that aldehyde dehydrogenase (ALDH) 1a1 is the major ALDH expressed in mouse liver and is an effective catalyst in metabolism of lipid aldehydes. Quantitative real-time polymerase chain reaction analysis revealed a Ϸ2.5-to 3-fold induction of the hepatic ALDH1A1 mRNA in mice administered either acrolein (5 mg/kg acrolein p.o.) or butylated hydroxylanisole (BHA) (0.45% in the diet) and of cytosolic NAD ϩ -dependent ALDH activity. We observed Ϸ2-fold increases in ALDH1A1 mRNA levels in both Nrf2(ϩ/ϩ) and Nrf2(Ϫ/Ϫ) mice treated with BHA compared with controls, suggesting that BHA-induced expression is independent of nuclear factor E2-related factor 2 (Nrf2). The levels of activator protein-1 (AP-1) mRNA and protein, as well as the amount of phosphorylated c-Jun were significantly increased in mouse liver or Hepa1c1c7 cells treated with either BHA or acrolein. With use of luciferase reporters containing the 5Ј-flanking sequence of Aldh1a1 (Ϫ1963/ϩ27), overexpression of c-Jun resulted in an Ϸ4-fold induction in luciferase activity, suggesting that c-Jun transactivates the Aldh1a1 promoter as a homodimer and not as a c-Jun/c-Fos heterodimer. Promoter deletion and mutagenesis analyses demonstrated that the AP-1 site at position Ϫ758 and possibly Ϫ1069 relative to the transcription start site was responsible for c-Jun-mediated transactivation. Electrophoretic mobility shift assay analysis with antibodies against c-Jun and c-Fos showed that c-Jun binds to the proximal AP-1 site at position Ϫ758 but not at Ϫ1069. Recruitment of c-Jun to this proximal AP-1 site by BHA was confirmed by chromatin immunoprecipitation analysis, indicating that recruitment of c-Jun to the mouse Aldh1a1 gene promoter results in increased transcription. This mode of regulation of an ALDH has not been described before.

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Section: Methodssupporting

confidence: 49%

“…Previous studies showed that Nrf2 induces the expression of cytoprotective genes by forming active heterodimers with AP-1 proteins, in particular c-Jun (Lee and Murray, 2010). We examined whether there is cross-talk between c-Jun and Nrf2 in Aldh1a1 transcriptional activation.…”

Section: Constitutivementioning

confidence: 99%

Activator Protein-1 Regulation of Murine Aldehyde Dehydrogenase 1a1

Makia

Amunom²,

Falkner³

et al. 2012

Mol Pharmacol

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“…Concerning the role of Nrf2 in the regulation of P450, mouse Cyp2a5 was the first reported case [12]. In addition, a recent study demonstrated that human CYP2J2 is regulated by Nrf2 [21]. We could provide evidence to put CYP2A6 into the short list of P450s that are regulated by Nrf2.…”

Section: Discussionmentioning

confidence: 88%

Human CYP2A6 is regulated by nuclear factor-erythroid 2 related factor 2

Yokota

Higashi

Fukami

et al. 2011

Biochemical Pharmacology

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Human CYP2A6 is responsible for the metabolism of nicotine and coumarin as well as the metabolic activation of tobacco-related nitrosamines. Earlier studies revealed that CYP2A6 activity was increased by dietary cadmium or cruciferous vegetables, but the underlying mechanisms remain to be clarified. In the present study, we investigated the possibility that Nrf2 might be involved in the regulation of CYP2A6. Real-time RT-PCR analysis revealed that the CYP2A6 mRNA level in human hepatocytes was significantly (P < 0.01, 1.4 fold) induced by 10 µM sulforaphane (SFN), a typical activator of Nrf2. A computer-based search identified three putative antioxidant response elements (AREs) in the 5'-flanking region of the CYP2A6 gene at positions -1212, -2444, and -3441, termed ARE1, ARE2, and ARE3, respectively. Electrophoretic mobility shift assays demonstrated that Nrf2 bound only to ARE1. Luciferase assays using HepG2 cells revealed that the overexpression of Nrf2 significantly increased the reporter activities of the constructs containing a 30-bp fragment that included ARE1. However, the activity of the construct containing the intact 5'-flanking region (-1 to -1395) including ARE1 was not increased by the overexpression of Nrf2. In contrast, when the reporter construct was injected into mice via the tail vein, the reporter activity in the liver was significantly (P < 0.05, 1.9 fold) increased by SFN (1 mg/head) administration. In conclusion, we found that human CYP2A6 is regulated via Nrf2, suggesting that CYP2A6 is induced under oxidative stress.

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“…It is important to note that Lee and Murray (2010) induced their cells with BHA for 72 hours compared with the 48 hours of this study. Further, they replenished the BHA in their cell media frequently during their induction (at 6, 12, 18, 24, and 48 hours), whereas BHA was replenished at 24 hours in this study.…”

Section: Discussionmentioning

confidence: 99%

“…A 48-hour induction period was chosen after preliminary studies indicated that significant cell death occurred at 72 hours. Lee and Murray (2010) reported BHA as a CYP2J2 inducer in HepG2 cells. Further work by Ma et al (2004) has shown that the mouse ortholog CYP2J5 is regulated by sex hormones in murine kidneys.…”

Section: Discussionmentioning

confidence: 99%

Activity, Inhibition, and Induction of Cytochrome P450 2J2 in Adult Human Primary Cardiomyocytes

et al. 2013

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Cytochrome P450 2J2 plays a significant role in the epoxidation of arachidonic acid to signaling molecules important in cardiovascular events. CYP2J2 also contributes to drug metabolism and is responsible for the intestinal clearance of ebastine. However, the interaction between arachidonic acid metabolism and drug metabolism in cardiac tissue, the main expression site of CYP2J2, has not been examined. Here we investigate an adult-derived human primary cardiac cell line as a suitable model to study metabolic drug interactions (inhibition and induction) of CYP2J2 in cardiac tissue. The primary human cardiomyocyte cell line demonstrated similar mRNA-expression profiles of P450 enzymes to adult human ventricular tissue. CYP2J2 was the dominant isozyme with minor contributions from CYP2D6 and CYP2E1. Both terfenadine and astemizole oxidation were observed in this cell line, whereas midazolam was not metabolized suggesting lack of CYP3A activity. Compared with recombinant CYP2J2, terfenadine was hydroxylated in cardiomyocytes at a similar K m value of 1.5 mM. The V max of terfenadine hydroxylation in recombinant enzyme was found to be 29.4 pmol/pmol P450 per minute and in the cells 6.0 pmol/pmol P450 per minute. CYP2J2 activity in the cell line was inhibited by danazol, astemizole, and ketoconazole in submicromolar range, but also by xenobiotics known to cause cardiac adverse effects. Of the 14 compounds tested for CYP2J2 induction, only rosiglitazone increased mRNA expression, by 1.8-fold. This cell model can be a useful in vitro model to investigate the role of CYP2J2-mediated drug metabolism, arachidonic acid metabolism, and their association to drug induced cardiotoxicity.

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Up-Regulation of Human CYP2J2 in HepG2 Cells by Butylated Hydroxyanisole Is Mediated by c-Jun and Nrf2

Abstract: Cytochrome P450 2J2 oxidizes arachidonic acid to a series of epoxyeicosatrienoic acid (EET) isomers in human tissues. EETs regulate numerous homeostatic processes, including cytoprotective and proliferative responses against injurious stresses.

Cited by 26 publications

References 34 publications

Activator Protein-1 Regulation of Murine Aldehyde Dehydrogenase 1a1

Activator Protein-1 Regulation of Murine Aldehyde Dehydrogenase 1a1

Human CYP2A6 is regulated by nuclear factor-erythroid 2 related factor 2

Activity, Inhibition, and Induction of Cytochrome P450 2J2 in Adult Human Primary Cardiomyocytes

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