Long-term effects of a standardized complex mixture of urban dust particulate on the metabolic activation of carcinogenic polycyclic aromatic hydrocarbons in human cells in culture.

Humans are exposed to complex mixtures of polycyclic aromatic hydrocarbons in the atmosphere. We examined the long term effects of a standard reference material (SRM) 1649a over time on the metabolic activation and DNA adduct formation by two carcinogenic PAHs, benzo[a]pyrene (BP) and dibenzo[a,l]pyrene (DBP) in the human mammary carcinoma derived cell line MCF-7. PAH-DNA adduct analysis, cytochrome P450 (CYP) enzyme activity, CYP1A1 and CYP1B1 protein expression were determined in cells treated with SRM 1649a alone or SRM 1649a with either BP or DBP for 24 to 120 h. Averaging over time, significantly higher levels of DNA adducts were observed in cells treated with BP or DBP alone than in co-treatments with SRM 1649a and BP or DBP. Ethoxyresorufin O-deethylase assay indicated a significant increase in activity in cells treated with BP alone and co-treated with SRM1649a in comparison to other treatment groups. Induction of CYP1A1 and CYP1B1 protein expression was observed by immunoblots in cells treated with BP alone or in co-treatments of SRM 1649a and BP or DBP. These data demonstrate the influence of the components of SRM 1649a in inhibiting the activation of BP or DBP by CYP enzymes in the formation of DNA adducts. It also suggests that the carcinogenic activity of PAH within a complex mixture may vary with composition and activation of the components present in the complex mixture.     

Sensitivity of Syrian golden hamster fetal lung cells to benzo[a]pyrene and other polycyclic hydrocarbons in vitro

Dose responses were compared of cultured fetal Syrian golden hamster lung cells (FSHL) to the toxic and transforming effects of benzo[a]pyrene (B[a]P), benzo[b]fluoranthene (B[b]F), benz[a]anthracene (B[a]a) indeno[1,2,3-c,d]pyrene (I[c,d]P), benzo[k]fluoranthene (B[k]F) and benzo[e]pyrene (B[e]P). Effort was first given to standardising the techniques for evaluating B[e]P dose-responses. These polycyclic aromatic hydrocarbons (PAH) were then tested at concentrations of up to 1 μg/ml, and only B[a]P showed clear cytotoxicity. The transforming effects of B[b]F, B[a]A and I[c,d]P at 1 μg/ml appeared comparable to those of B[a]P at 0.05 μg/ml.     

Importance of CYP1A1 and CYP1B1 in bioactivation of benzo[a]pyrene in human lung cell lines

Polycyclic aromatic hydrocarbons are ubiquitous environmental pollutants classified as carcinogens in humans and rodents. The cytochromes P4501A1 and 1B1 have both shown capacity to carry out bioactivation of the prototype PAH, benzo[a]pyrene (B[a]P) to its ultimate carcinogenic B[a]P-diol-epoxide-I-1 form. The part played by each enzyme in human lung cells, however, has not been clarified. To get further insight into their individual role in the metabolic activation of B[a]P, RNA-interference was used to down-regulate CYP1A1 and/or CYP1B1 gene expression in the human lung cell lines BEP2D and NCIH2009. Fluorescence-HPLC analysis revealed that formation of B[a]P-tetrol-I-1 (hydrolyzed form of the corresponding diol-epoxide) was dependent primarily on CYP1A1. In cells without down-regulation of CYP1A1, the B[a]P-tetrol-I-1 was the major tested isomer formed. In contrast, the B[a]P-cis- and trans-7,8-dihydrodiol isomers were readily formed in cells expressing high levels of either CYP-gene. Simultaneous down-regulation of CYP1A1 and CYP1B1 mRNA resulted in low levels of metabolites overall. Residual unmetabolized B[a]P levels followed the expression of CYP1A1 in an inverse manner. In conclusion, these results indicate a major role of CYP1A1 in the bioactivation of B[a]P to carcinogenic B[a]P-diol-epoxides and in overall metabolism of B[a]P in human lung cell lines. In contrast, both CYP1A1 and CYP1B1 contribute significantly to the formation of the B[a]P-cis- and trans-7,8-dihydrodiol isomers.     

Albumin and hemoglobin adducts of benzo[a]pyrene in humans—Analytical methods,exposure assessment,and recommendations for future directions

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous in our environment and can cause cancer. Exposure to PAHs can be assessed by protein adduct dosimetry using benzo[a]pyrene (B[a]P) as a model compound. We present an overview of analytical methods to detect B[a]P- derived protein adducts in humans, their uses in exposure assessment, and recommendations for future research. Two major methodologies, enzyme-linked immunosorbent assay (ELISA) and chemical-specific assays, could be traced in the literature but there remains limitations with both assays. ELISA is nonspecific due to cross-reactivity of the antibody with other PAHs and results are better interpreted in terms of PAH exposure. ELISA is unable to distinguish between exposed and nonexposed persons in the majority of studies. Adduct concentrations are higher by several orders of magnitude compared to those determined by chemical-specific methods. The latter methods mostly analyzed protein adducts derived by (+)-anti-B[a]P-diol epoxide. For this purpose, gas or liquid chromatography in combination with mass spectrometry or fluorescence detection were used. However, the prevalence of positive samples remained low when chemical- specific assays were used mainly due to the lack of sensitivity. Overall, data on B[a]P-derived protein adducts in humans remain inconclusive. Future research should focus on the development and standardization of a sensitive and specific method for B[a]P-derived protein adducts prior to its use in field studies. Finally, exposures of B[a]P at the workplace and via diet, a major route of exposure of the general population, can be studied. The results will contribute to the understanding of B[a]P-induced cancer and will allow for health preventive measures.     

Quantitative microspectrofluorimetry study of the “blocking effect” of 6-aminochrysene on benzo[a]pyrene metabolism using single living cells

By microspectrofluorimetry on single living cells (murine fibroblasts 3T3), we have obtained monoexponential decreases of fluorescence intensity for benzo[a]pyrene (B[a]P) and 6-aminochrysene (6a-chrysene) metabolism. These kinetics are characteristics of B[a]P and 6a-chrysene metabolism and histograms can be drawn from the rate constants. We have studied the influence of 6a-chrysene on B[a]P metabolism. Using different methods of incubation, it has been observed that the presence of 6a-chrysene leads to modifications of the histogram profiles during B[a]P metabolism. Polycyclic aromatic hydrocarbons (PAH) are used to induce B[a]P metabolism. Whatever the experimental conditions we never detected such a phenomenon with 6a-chrysene. On the contrary we have observed an inhibition of B[a]P metabolism by 6a-chrysese, which can reach 80% of the aryl hydrocarbon hydroxylase (AHH) activity when 6a-chrysene remains constant in the cells. Compared with the results previously observed “in vitro” (which presented 50% mean inhibition) we show that inhibiyion acts in an all-or-nothing mechanism at the cellular level.     

Benzo[a]pyrene at human blood equivalent level induces human lung epithelial cell invasion and migration via aryl hydrocarbon receptor signaling

Benzo[a]pyrene (B[a]P), a typical carcinogenic polycyclic aromatic hydrocarbon, exists worldwide in vehicle exhaust, cigarette smoke and other polluted environments. Recent studies have demonstrated a strong association between B[a]P and lung cancer. However, whether B[a]P at human blood equivalent level can promote epithelial-mesenchymal transition (EMT), a crucial molecular event during cell malignant transformation, remains unclear. Besides, whether B[a]P facilitates this progress via aryl hydrocarbon receptor (AhR) signaling pathway also lacks scientific evidence. In our study, the transwell assay showed that 5 μg/L of B[a]P promoted BEAS-2B cell invasion and migration. In addition, the mRNA and protein expression levels of AhR and its target genes involved in B[a]P metabolism, such as AhR nuclear translocator, heat shock protein 90 and CYP1A1, were significantly increased by B[a]P exposure. Moreover, the mRNA expression levels of downstream regulatory factors related to both AhR signaling pathway and EMT, such as NRF2, K-RAS and hypoxia-inducible factor 1-alpha, were significantly increased. Furthermore, the expression level of the epithelial marker E-cadherin was significantly downregulated, while the mRNA expression of mesenchymal phenotype markers, N-cadherin, fibronectin and vimentin, were significantly upregulated. Notably, the above changes induced by B[a]P were significantly attenuated or even stopped by resveratrol (RSV), a natural phenol, also an AhR inhibitor, when the AhR signaling pathway was inhibited by RSV, demonstrating the regulatory role of AhR signaling pathway in B[a]P-induced EMT. In conclusion, B[a]P at the human blood equivalent level induces BEAS-2B cell invasion and migration through the AhR signaling pathway.     

Transformation of BHK 21/CL 13 cells by various polycyclic aromatic hydrocarbons using the method of styles

Nine polycyclic aromatic hydrocarbons (PAHs) were investigated by the cell-transformation assay method of Styles. Benzo(a)pyrene [B(a)P], chrysene (CH), 3-methylcholanthrene (3-MC), benz(a)anthracene (BA), benzo(b)fluoranthene [B(b)F], and dibenz(a,h)anthracene (DBA) were tested, including liver homogenate, and showed dose-effect relationships and a more than 2-fold increase of transformation rates at LC₅₀. Due to variations of the test method our results differed quantitatively from the data published by Purchase and Styles. Discrimination between the known carcinogens listed above and the noncarcinogens, phenanthrene (PA) and anthracene (AC), lacking a dose-effect relationship was, however, possible. Benzo(e)pyrene [B(e)P] was regarded as positive although producing only a 2-fold increase in the number of transformed colonies.     

Multidrug resistance protein (MRP) 4 attenuates benzo[a]pyrene-mediated DNA-adduct formation in human bronchoalveolar H358 cells

Multi-drug resistance protein (MRP) 4, an ATP-binding cassette (ABC) transporter, has broad substrate specificity. It facilitates the transport of bile salt conjugates, conjugated steroids, nucleoside analogs, eicosanoids, and cardiovascular drugs. Recent studies in liver carcinoma cells and hepatocytes showed that MRP4 expression is regulated by the aryl hydrocarbon receptor (AhR) and nuclear factor E2-related factor 2 (Nrf2). The AhR has particular importance in the lung and is most commonly associated with the up-regulation of cytochrome P-450 (CYP)-mediated metabolism of benzo[a]pyrene (B[a]P) to reactive intermediates. Treatment of H358, human bronchoalveolar, cells with 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) or (−)-benzo[a]pyrene-7,8-dihydro-7,8-diol (B[a]P-7,8-dihydrodiol), the proximate carcinogen of B[a]P, revealed that MRP4 expression was increased compared to control. This suggested that MRP4 expression might contribute to the paradoxical decrease in (+)-7,8-dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene-2′-deoxyguanosine ((+)-anti-trans-B[a]PDE-dGuo) DNA-adducts observed in TCDD-treated H358 cells. We have now found that decreased MRP4 expression induced by a short hairpin RNA (shRNA), or chemical inhibition with probenecid, increased (+)-anti-trans-B[a]PDE-dGuo formation in cells treated with (−)-B[a]P-7,8-dihydrodiol, but not the ultimate carcinogen (+)-anti-trans-B[a]PDE. Thus, up-regulation of MRP4 increased cellular efflux of (−)-B[a]P-7,8-dihydrodiol, which attenuated DNA-adduct formation. This is the first report identifying a specific MRP efflux transporter that decreases DNA damage arising from an environmental carcinogen.     

In vitro metabolism of benzo[a]pyrene and dibenzo[def,p]chrysene in rodent and human hepatic microsomes

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous and often carcinogenic contaminants released into the environment during natural and anthropogenic combustion processes. Benzo[a]pyrene (B[a]P) is the prototypical carcinogenic PAH, and dibenzo[def,p]chrysene (DBC) is a less prevalent, but highly potent transplacental carcinogenic PAH. Both are metabolically activated by isoforms of the cytochrome P450 enzyme superfamily to form reactive carcinogenic and cytotoxic metabolites. Metabolism of B[a]P and DBC was studied in hepatic microsomes of male Sprague-Dawley rats, naïve and pregnant female B6129SF1/J mice, and female humans, corresponding to available pharmacokinetic data. Michaelis–Menten saturation kinetic parameters including maximum rates of metabolism (V_MAX, nmol/min/mg microsomal protein), affinity constants (K_M, μM), and rates of intrinsic clearance (CL_INT, ml/min/kg body weight) were calculated from substrate depletion data. CL_INT was also estimated from substrate depletion data using the alternative in vitro half-life method. V_MAX and CL_INT were higher for B[a]P than DBC, regardless of species. Clearance for both B[a]P and DBC was highest in naïve female mice and lowest in female humans. Clearance rates of B[a]P and DBC in male rat were more similar to female human than to female mice. Clearance of DBC in liver microsomes from pregnant mice was reduced compared to naïve mice, consistent with reduced active P450 protein levels and elevated tissue concentrations and residence times for DBC observed in previous in vivo pharmacokinetic studies. These findings suggest that rats are a more appropriate model organism for human PAH metabolism, and that pregnancy's effects on metabolism should be further explored.     

Urban dust particulate matter alters PAH-induced carcinogenesis by inhibition of CYP1A1 and CYP1B1.

The polycyclic aromatic hydrocarbons (PAHs) benzo[a]pyrene (B[a]P) and dibenzo[a,l]pyrene (DB[a,l]P) are well-studied environmental carcinogens, however, their potency within a complex mixture is uncertain. We investigated the influence of urban dust particulate matter (UDPM) on the bioactivation and tumor initiation of B[a]P and DB[a,l]P in an initiation-promotion tumorigenesis model. SENCAR mice were treated topically with UDPM or in combination with B[a]P or DB[a,l]P, followed by weekly application of the promoter 12-O-tetradecanoylphorbol-13 acetate. UDPM exhibited weak tumor-initiating activity but significantly delayed the onset of B[a]P-induced tumor initiation by two-fold. When cotreated with UDPM, DB[a,l]P-treated animals displayed no significant difference in tumor-initiating activity, compared with DB[a,l]P alone. Tumor initiation correlated with PAH-DNA adducts, as detected by (33)P-postlabeling and reversed-phase high-performance liquid chromatography. Induction of cytochrome P450 (CYP)1A1 and 1B1 proteins was also detected following UDPM treatment or cotreatment with B[a]P or DB[a,l]P, indicating PAH bioactivation. Further genotoxicity analyses by the comet assay revealed that cotreatment of UDPM plus B[a]P or DB[a,l]P resulted in increased DNA strand breaks, compared with PAH treatment alone. The metabolizing activities of CYP1A1 and CYP1B1, as measured by the 7-ethoxyresorufin O-deethylation (EROD) assay, revealed that UDPM noncompetitively inhibited CYP1A1 and CYP1B1 EROD activity in a dose-dependent manner. Overall, these data suggest that components within complex mixtures can alter PAH-induced carcinogenesis by inhibiting CYP bioactivation and influence other genotoxic effects, such as oxidative DNA damage. These data further suggest that in addition to the levels of potent PAH, the effects of other mixture components must be considered when predicting human cancer risk.     

Aldehyde dehydrogenase 1A1 up-regulates stem cell markers in benzo[a]pyrene-induced malignant transformation of BEAS-2B cells

Recently, Aldehyde dehydrogenase 1A1 (ALDH1A1) has been proposed to be a common marker of cancer stem cells and can be induced by benzo[a]pyrene (B[a]P) exposure. However, the underlying mechanism of how ALDH1A1 contributes to B[a]P-induced carcinogenesis in human bronchial epithelial cells remains unclear. Here, we found that B[a]P up-regulated expression levels of stem cell markers (ABCG2, SOX2, c-Myc and Klf4), epithelial-mesenchymal transition (EMT) associated genes (SNAIL1, ZEB1, TWIST and β-CATENIN) and cancer-related long non-coding RNAs (lncRNAs; HOTAIR and MALAT-1) in malignant B[a]P-transformed human bronchial epithelial cells (BEAS-2B-T cells), and these up-regulations were dependent on increased expression of ALDH1A1. The inhibition of endogenous ALDH1A1 expression down-regulated expression levels of stem cell markers and reversed the malignant phenotype as well as reduced the chemoresistance of BEAS-2B-T cells. In contrast, the overexpression of ALDH1A1 in BEAS-2B cells increased the expression of stem cell markers, facilitated cell transformation, promoted migratory ability and enhanced the drug resistance of BEAS-2B cells. Overall, our data indicates that ALDH1A1 promotes a stemness phenotype and plays a critical role in the BEAS-2B cell malignant transformation induced by B[a]P.     

A Search for the Plant Ingredients that Protect Cells from Air Pollutants and Benz[a]pyrene Phototoxicity

A diversity of antioxidants and plant ingredients were examined for their protective effect in cultured Balb/c 3T3 cells against ultraviolet A (UVA)-induced cytotoxicities of extracted air pollutants and benz[a]pyrene (B[a]P) in an effort to find effective protectors against the phototoxicity of air pollutants and B[a]P. As has been observed for B[a]P phototoxicity, air pollutants themselves and those previously exposed to UVA light in the absence of cells exhibited faintly weak cytotoxicity, but the toxicity was markedly elevated when they were exposed to UVA light concomitantly with cells. The B[a]P phototoxicity was not eliminated by well-known antioxidants but was markedly diminished by diversity of plant ingredients. Among the plant ingredients tested in the current study, morin, naringin, and quercetin were found to be desirable protectors against B[a]P phototoxicity.     

Surface Marker-Defined Head Kidney Granulocytes and B Lymphocytes of Rainbow Trout Express Benzo[a]pyrene-Inducible Cytochrome P4501A Protein

Polycyclic aromatic hydrocarbons (PAHs) such as benzo[a]pyrene(BaP) are immunotoxic to fish. Metabolism of PAHs in immunecells has been implicated in PAH immunotoxicity in mammals,but for fish the presence of metabolic enzymes in immune cellsis less clear. The objective of this study was to examine localizationand induction of the BaP-metabolizing biotransformation enzyme,cytochrome P4501A (CYP1A), in head kidney immune cells of rainbowtrout (Oncorhynchus mykiss). In the first step, we measuredinduction of CYP1A-dependent 7-ethoxyresorufin-O-deethylase(EROD) activity and CYP1A protein in head kidney of rainbowtrout treated with a single intraperitoneal (ip) injection of25 mg BaP/kg body weight. From days 3 to 10 postinjection, theBaP treatment led to a significant elevation of EROD and CYP1Aprotein in head kidney and liver, with CYP1A expression levelsin the head kidney being much lower than in the liver. Next,we examined the cellular localization of CYP1A protein in thehead kidney cell types: vascular endothelial, endocrine andlymphoid cells. CYP1A immunoreactivity was detectable only inBaP-treated trout, where it was localized in endothelial andlymphoid cells. Finally, we aimed to clarify which of the hematopoieticcell types possess CYP1A protein. Using double immunostainingfor CYP1A and surface markers of rainbow trout immune cells,we identified B lymphocytes and granulocytes expressing inducibleCYP1A protein and being the likely sites of BaP metabolism inthe head kidney.     

Influence of red blood cells,serum albumin,and serum lipoproteins on the clearance of benzo[a]pyrene by isolated livers of 3-methylcholanthrene-treated rats

Red blood cells, serum albumin, and serum lipoproteins transport benzo[a]pyrene and other xenobiotic compounds in the circulation. The distribution of benzo[a]pyrene and its metabolites among these blood components was examined, and the effect of their presence in the perfusion medium on the ability of isolated livers from 3-methylcholanthrene-pretreated rats to clear circulating benzo[a]pyrene was determined. A large fraction (45%) of the benzo[a]pyrene in rat blood was associated with the serum lipoproteins. However, only 8% of the benzo[a]pyrene metabolites was associated with this component. Forty to forty-five percent of each was associated with red blood cells. Benzo[a]pyrene clearance by isolated rat livers was 1.8 ± 0.2 ml/min when the medium contained only red blood cells and buffer. Addition of serum lipoproteins or serum albumin increased benzo[a]pyrene clearance to 5.1 ± 0.5 or 8.5 ± 0.9 ml/min respectively. Appearance of benzo[a]pyrene metabolites in perfusion medium and bile was similarly altered by the changes in medium composition. These results indicate that the clearance of benzo[a]pyrene by rat liver depends on the composition of the medium perfusing the organ and suggest that alterations in blood components in vivo may influence the metabolic disposition of this carcinogen.     

Sidestream tobacco smoke as the main predictor of exposure to polycyclic aromatic hydrocarbons

Polycyclic aromatic hydrocarbons (PAHs) were measured in mainstream and sidestream tobacco smoke from 14 commercial brands of cigarettes purchased in Italy during 2001-2002. The PAHs detected in smoke and analysed with HPLC and a fluorimetric detector were: fluoranthene, pyrene, benzo[a]anthracene, chrysene, benzo[a]pyrene, benzo[b] fluoranthene, benzo[k] fluoranthene, benzo[g,h,i]perylene and dibenzo[a,h]anthracene. The PAH levels in mainstream smoke from different cigarette brands obtained using an official smoking machine varied by about threefold (from 118 to 374 ng per cigarette for total PAHs and from 23.5 to 100 ng per cigarette for carcinogenic PAHs). Total PAH levels in mainstream smoke were correlated with tar content (r = 0.615, P < 0.05, n = 14). Total PAH content in sidestream smoke, measured by collection of all the smoke produced by a lit cigarette in a glass chamber, was about tenfold higher compared with mainstream smoke. The PAH content in sidestream smoke was relatively uniform (2.3-3.9 and 0.49-1.21 micro g per cigarette for total and carcinogenic PAHs, respectively) and was not correlated with tar content. These results indicate that cigarette manufacturing and filter characteristics influence the PAH composition of mainstream smoke, but have no effect on the PAH content in sidestream smoke, which is the main determinant of smokers' and non-smokers' exposure to PAHs in environmental tobacco smoke.     

Induction of carbonyl reductase 1 (CBR1) expression in human lung tissues and lung cancer cells by the cigarette smoke constituent benzo[a]pyrene

Carbonyl reductase 1 (CBR1) reduces various xenobiotic carbonyl substrates to corresponding alcohol metabolites. Here we demonstrated that benzo[a]pyrene (B[a]P), a potent pro-carcinogen and predominant polycyclic aromatic hydrocarbon (PAH) compound in cigarette smoke and air pollutants, upregulates CBR1 gene expression in vitro and in vivo, and that a proximal xenobiotic response element (XRE) motif (₋₁₂₂XRE) mediates the induction effect of B[a]P. First, we observed 46% and 50% increases in CBR1 mRNA and CBR1 protein levels, respectively, in human lung tissue samples from smokers compared to never-smokers. Second, we detected 3.0-fold (p < 0.0001) induction of CBR1 mRNA and 1.5-fold (p < 0.01) induction of CBR1 protein levels in cells of the human lung cancer cell line A549 incubated with 2.5 μM B[a]P for 24 h. Third, results from experiments with CBR1 promoter constructs indicated that a proximal XRE motif (₋₁₂₂XRE) mediates induction of reporter activity in response to B[a]P. Furthermore, we detected enhanced nuclear translocation of aryl hydrocarbon receptor (AhR) following B[a]P exposure in A549 cells. Finally, we demonstrated increased binding of specific protein complexes to ₋₁₂₂XRE in nuclear extracts from B[a]P-treated cells and the presence of the AhR/Arnt complex in the specific nuclear protein ₋₁₂₂XRE complexes.     

Aryl hydrocarbon receptor-independent up-regulation of intracellular calcium concentration by environmental polycyclic aromatic hydrocarbons in human endothelial HMEC-1 cells

Polycyclic aromatic hydrocarbons (PAHs) such as benzo(a)pyrene (B(a)P) constitute a major family of widely‐distributed environmental toxic contaminants, known as potent ligands of the aryl hydrocarbon receptor (AhR). B(a)P has been recently shown to trigger an early and transient increase of intracellular calcium concentration ([Ca²⁺]_i), involved in AhR‐related up‐regulation of target genes by B(a)P. This study was designed to determine whether AhR may play a role in [Ca²⁺]_i induction provoked by B(a)P. We demonstrated that, in addition to B(a)P, various PAHs, including pyrene and benzo(e)pyrene, known to not or only very poorly interact with AhR, similarly up‐regulated [Ca²⁺]_i in human endothelial HMEC‐1 cells. Moreover, α‐naphthoflavone, a flavonoïd antagonist of AhR, was also able to induce [Ca²⁺]_i. Knocking‐down AhR expression in HMEC‐1 cells through transfection of siRNAs, was finally demonstrated to not prevent B(a)P‐mediated induction of [Ca²⁺]_i, whereas it efficiently counteracted B(a)P‐mediated induction of the referent AhR target gene cytochrome P‐450 1B1. Taken together, these data demonstrate that environmental PAHs trigger [Ca²⁺]_i induction in an AhR‐independent manner. © 2011 Wiley Periodicals, Inc. Environ Toxicol, 2012.     

Preliminary physiologically based pharmacokinetic models for benzo[a]pyrene and dibenzo[def,p]chrysene in rodents

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental contaminants generated as byproducts of natural and anthropogenic combustion processes. Despite significant public health concern, physiologically based pharmacokinetic (PBPK) modeling efforts for PAHs have so far been limited to naphthalene, plus simpler PK models for pyrene, nitropyrene, and benzo[a]pyrene (B[a]P). The dearth of published models is due in part to the high lipophilicity, low volatility, and myriad metabolic pathways for PAHs, all of which present analytical and experimental challenges. Our research efforts have focused upon experimental approaches and initial development of PBPK models for the prototypic PAH, B[a]P, and the more potent, albeit less studied transplacental carcinogen, dibenzo[def,p]chrysene (DBC). For both compounds, model compartments included arterial and venous blood, flow limited lung, liver, richly perfused and poorly perfused tissues, diffusion limited fat, and a two compartment theoretical gut (for oral exposures). Hepatic and pulmonary metabolism was described for both compounds, as were fractional binding in blood and fecal clearance. Partition coefficients for parent PAH along with their diol and tetraol metabolites were estimated using published algorithms and verified experimentally for the hydroxylated metabolites. The preliminary PBPK models were able to describe many, but not all, of the available data sets, comprising multiple routes of exposure (oral, intravenous) and nominal doses spanning several orders of magnitude. Supported by Award Number P42 ES016465 from the National Institute of Environmental Health Sciences.