Polycyclic aromatic hydrocarbons stimulate human CYP3A4 promoter activity via PXR

Luckert, Claudia; Ehlers, Anke; Buhrke, Thorsten; Seidel, Albrecht; Lampen, Alfonso; Hessel, Stefanie

doi:10.1016/j.toxlet.2013.06.243

Cited by 83 publications

(68 citation statements)

References 35 publications

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“…Proteins were separated on 12 % SDS polyacrylamide gels (20 µg of protein per lane) and blotted onto nitrocellulose membranes as described by (Luckert et al 2013). The blots were processed by using the following antibodies: anti-GST3/GST pi antibody [EPR8263] (ab138491; Abcam; dilution 1:10,000); anti-cyclophilin B antibody [EPR12703(B)]-loading control (ab178397; Abcam; dilution 1:10,000); goat anti-rabbit IgG-HRP (Sc2004; Santa Cruz Biotechnology; dilution 1:10,000).…”

Section: Western Blottingmentioning

confidence: 99%

“…The highcapacity cDNA reverse transcription kit (Applied Biosystems/Life Technologies GmbH, Darmstadt, Germany) was employed to reverse-transcribe RNA samples. Subsequently, cDNA samples were subjected to qPCR analysis by using the Maxima SYBR Green/Rox qPCR Master Mix (Thermo Fisher Scientific, Waltham, MA, USA) and a Stratagene MX3005P real-time PCR cycler (Agilent Technologies, Santa Clara, CA, USA) according to the protocol by (Luckert et al 2013). Gene expression was calculated according to the C t method by using the expression values of the housekeeping gene cyclophilin B (Ppib) for normalization.…”

Section: Quantitative Real-time Rt-pcr Analysismentioning

confidence: 99%

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Proteomic analysis of 3-MCPD and 3-MCPD dipalmitate-induced toxicity in rat kidney

et al. 2015

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3-Chloropropane-1,2-diol (3-MCPD) and its fatty acid esters are formed during thermal treatment of fat-containing foodstuff in the presence of salt. Toxicological studies indicate a carcinogenic potential of 3-MCPD, pointing to the kidney as the main target organ. It is assumed that the toxicological property of 3-MCPD esters is constituted by the release of 3-MCPD during digestion. In a repeated-dose 28-day oral toxicity study using Wistar rats, animals were treated with equimolar doses of either 3-MCPD (10 mg/kg body weight) or 3-MCPD dipalmitate (53 mg/kg body weight). A lower dose of 3-MCPD dipalmitate (13.3 mg/kg body weight) was also applied. No histopathologically visible toxicity was observed in the study. To address molecular mechanisms leading to toxicity of 3-MCPD and its esters, kidney samples were analyzed by a comparative, two-dimensional gel electrophoresis/mass spectrometry proteomic approach. After either 3-MCPD or 3-MCPD dipalmitate treatment, alterations in proteins related to various metabolic pathways, including carbohydrate, amino acid, and fatty acid metabolism, were detected. These findings confirm and complement previous data on the inhibition of glucose metabolism by 3-MCPD. Altogether, broad overlap of 3-MCPD- and 3-MCPD dipalmitate-induced proteomic changes was observed. Further analyses revealed that the observed induction of glutathione S-transferase pi 1 (Gstp1) occurred at the transcriptional level and was not related to nuclear factor (erythroid-derived 2)-like 2 activation. Overall, the results indicate common mechanisms of toxicity for 3-MCPD and its dipalmitate ester. Furthermore, data suggest Gstp1 as a sensitive marker for early 3-MCPD-induced effects in rat kidney.

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Section: Western Blottingmentioning

confidence: 99%

Section: Quantitative Real-time Rt-pcr Analysismentioning

confidence: 99%

Proteomic analysis of 3-MCPD and 3-MCPD dipalmitate-induced toxicity in rat kidney

et al. 2015

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“…We previously reported that indirubin, a ligand of AhR, induces CYP3A4 through PXR (Kumagai et al, 2012;Kumagai et al, 2016) and other group also reported PAHs induce CYP3A4 (Luckert et al, 2013). In addition, indirubin showed a similar profile of transcriptional activation of CYP3A4 to that of TCDD and DBA (data not shown) in HepG2 cells.…”

Section: Discussionmentioning

confidence: 64%

CYP3A4 induction mechanism of polycyclic aromatic hydrocarbons differs from that of rifampicin in PXR binding element

Aratsu

Odagiri

Shoji

et al. 2017

Fundam. Toxicol. Sci.

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-CYP3A4 is an important drug-metabolizing enzyme induced by various compounds causing drug-drug interactions. However, the molecular mechanism of CYP3A4 induction is not completely understood. CYP3A4 induction is caused by pregnane X receptor (PXR) through binding to some PXR binding elements. These elements comprise an everted repeat separated by six nucleotides in the promoter region and distal nuclear receptor binding element 1 (dNR-1) as well as the essential distal nuclear receptor binding element for CYP3A4 induction (eNR3A4) in the enhancer region of the CYP3A4 gene. Recently, we found that polycyclic aromatic hydrocarbons including anthracene induce CYP3A4 in HepG2 cells with a different induction profile from that of rifampicin (RF), a typical PXR ligand. When a CYP3A4 reporter plasmid in which the eNR3A4 DNA fragment binds directly to the CYP3A4 promoter (-362 bases) was evaluated in a reporter assay, dibenz [a,h]anthracene (DBA) induced reporter activity, while RF did not. To be induced reporter activity by RF, more 14 nucleotides 5′ upstream of the eNR3A4 (rifampicin eNR3A4: reNR3A4) DNA fragment were required. However, eNR3A4 and reNR3A4 did not respond to recombinant PXR without dNR-1. These results suggest that eNR3A4 and reNR3A4 are necessary for CYP3A4 induction by DBA and RF, respectively, and that dNR-1 is indispensable for full induction through PXR.

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“…Gene expression of CYP3A27 has been reported to be regulated by pregnane X receptor (PXR) in rainbow trout hepatocytes (Wassmur et al 2010). Since PAHs activate the PXR signaling pathway in humans (Luckert et al 2013), it could be speculated that oil PAHs induce O. mykiss CYP3A27 expression through PXR. However, there are few studies on the nature of PXR ligands in fish (Ekins et al 2008, Krasowski et al 2011, Bainy et al 2013 and their results show differences in ligand affinity between fish and mammalian PXRs, and also among fish species.…”

Section: Discussionmentioning

confidence: 99%

“…The mRNA of CYP3A27 is 55 % identical to human CYP3A4, and their encoded proteins (LMC5-like) have structural and functional similarities (Miranda et al 1991). Through pregnane X receptor (PXR) signaling pathway, some polycyclic aromatic hydrocarbons induce human CYP3A4 (Luckert et al 2013). Similarly, aromatic hydrocarbons could act as endocrine-disrupting chemicals in O. mykiss by inducing CYP3A27 mRNA expression, possibly affecting fish reproduction.…”

Section: Introductionmentioning

confidence: 99%

CYPS MOLECULAR BIOMARKERS IN RAINBOW TROUT Oncorhynchus mykiss TO ASSESS OIL CONTAMINATION IN A PATAGONIAN STREAM

Leggieri

Anna²,

Cerón³

et al. 2017

Rev. Int. Contam. Ambie.

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Cytochrome P450 (CYP) gene expressions were studied by semi-quantitative polymerase chain reaction (PCR) to assess oil pollution using liver and gills of juvenile rainbow trout Oncorhynchus mykiss sampled from La Mina stream (Bariloche, Argentina), up and downstream of a discharge of petroleum spill from an unproductive wellbore. Data was complemented with laboratory exposure to water accommodated fraction (WAF) from the oil spill. Oil exposure induced gill and liver CYP1A transcription (1.2-1.4-fold), which was used as reference biomarker. In both organs, CYP2K1 transcript was induced after 5 % WAF exposure, 24 h (1.8-2.0-fold), and it was down-regulated in more extended time (1.2-1.5-fold). CYP2M1 transcript levels were down-regulated in liver of impacted wild trout and in trout exposed to 5 % WAF during 96 h (1.5-1.7-fold). CYP2K1 and CYP2M1 are involved in the metabolism of fatty acids and estrogenic hormones, and its down-regulation would indicate metabolic alterations. CYP3A27 levels were only induced in liver of wild trout (1.6-fold), possibly related to crosstalk between aryl hydrocarbon receptor and pregnane X receptor signaling pathway. While CYP1A mRNA expression appears to be a more robust biomarker for detecting the effects of crude oil under different dose and exposure time, the expression of CYP2K1, CYP2M1 and CYP3A27 mRNAs would indicate different time exposure to oil pollution, particularly under high oil concentration in water. Hence, patterns of multiple CYP genes expression in rainbow trout gills and liver could be especially valuable for monitoring programs of crude oil pollution and remediation actions on Patagonian freshwater ecosystems. L.R. Leggieri et al. 682Palabras clave: CYP450, derrame de petróleo, peces, arroyo contaminado, Patagonia RESUMEN Se estudió la expresión génica de citocromos P450 (CYP) mediante reacción semicuantitativa en cadena de la polimerasa (PCR, por sus siglas en inglés), en muestras de hígado y branquias de trucha arcoíris Oncorhynchus mykiss juveniles capturadas en un arroyo que es afectado por un derrame de petróleo. El estudio se completó con exposiciones in vivo a la fracción soluble del petróleo del vertedero (WAF). El CYP1A, usado como biomarcador de referencia, fue inducido por el petróleo (1.2-1.4-veces) en branquias e hígado. En ambos órganos, el CYP2K1 fue inducido después de 24 h con 5 % WAF (1.8-2.0-veces), y subregulado en tiempos más extensos (1.2-1.5-veces). CYP2M1 fue subregulado en el hígado de truchas contaminadas y de aquellas expuestas a 96 h con 5 % (1.5-1.7-veces). Dado que CYP2K1 y CYP2M1 participan del metabolismo de ácidos grasos y de hormonas estrogénicas, la subregulación indicaría alteraciones en estos metabolismos. CYP3A27 fue inducido sólo en el hígado de las truchas silvestres afectadas por petróleo, posiblemente por efectos cruzados entre las señalizaciones de los receptores de hidrocarburos de arilo y X de pregnano. Mientras que la expresión de CYP1A mRNA pareciera ser un biomarcador más robusto para detectar los efectos d...

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Polycyclic aromatic hydrocarbons stimulate human CYP3A4 promoter activity via PXR

Cited by 83 publications

References 35 publications

Proteomic analysis of 3-MCPD and 3-MCPD dipalmitate-induced toxicity in rat kidney

Proteomic analysis of 3-MCPD and 3-MCPD dipalmitate-induced toxicity in rat kidney

CYP3A4 induction mechanism of polycyclic aromatic hydrocarbons differs from that of rifampicin in PXR binding element

CYPS MOLECULAR BIOMARKERS IN RAINBOW TROUT Oncorhynchus mykiss TO ASSESS OIL CONTAMINATION IN A PATAGONIAN STREAM

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