Polycyclic aromatic hydrocarbons (PAHs) are widespread environmental contaminants that interact in a complex manner with both the aryl hydrocarbon receptor (AhR) and estrogen receptors (ER). Their potential endocrine-disrupting activities may depend on both inhibitory AhR-ER cross-talk and on AhR-dependent metabolic production of estrogenic PAH metabolites. Here, we analyzed the impact of AhR on estrogen-like effects of PAHs, such as benzo[a]pyrene (BaP), in particular, on control of cell cycle progression/cell proliferation. Using AhR knockout variant of estrogen-sensitive human breast cancer MCF-7 cells (MCF-7 AhRKO cells), we observed that the AhR-dependent control of cytochrome P450 family 1 (CYP1) expression played a major role in formation of estrogenic BaP metabolites, most notably 3-OH-BaP, which contributed to the ER-dependent induction of cell cycle progression/cell proliferation. Both BaP metabolism and the BaP-induced S-phase transition/cell proliferation were inhibited in MCF-7 AhRKO cells, whereas these cells remained sensitive towards both endogenous estrogen 17β-estradiol or hydroxylated BaP metabolites. BaP was found to increase the activity of ER-dependent luciferase reporter gene in wild-type MCF-7 cells; however, unlike its hydroxylated metabolite, BaP failed to stimulate luciferase activity in MCF-7 AhRKO cells. Similarly, estrogen-like effects of other known estrogenic PAHs, such as benz[a]anthracene or 3-methylcholanthrene, were diminished in MCF-7 AhRKO cells. Ectopic expression of human CYP1A1 and CYP1B1 enzymes partly restored both BaP metabolism and its effects on cell proliferation. Taken together, our data suggest that the AhR-dependent metabolism of PAHs contributes significantly to the impact of PAHs on cell proliferation in estrogen-sensitive cells.

Department of Chemistry and Toxicology Veterinary Research Institute 62100 Brno Czech Republic

Department of Cytokinetics Institute of Biophysics of the Czech Academy of Sciences 61265 Brno Czech Republic

Department of Experimental Biology Faculty of Science Masaryk University 61137 Brno Czech Republic

Department of Nutrition Institute of Basic Medical Sciences University of Oslo 0372 Oslo Norway

Genomic Medicine and Environmental Toxicology Department Instituto de Investigaciones Biomédicas Universidad Nacional Autónoma de México C U 04510 Mexico City Mexico

Zobrazit více v PubMed

Abdelrahim M., Ariazi E., Kim K., Khan S., Barhoumi R., Burghardt R., Liu S., Hill D., Finnell R., Wlodarczyk B. (2006). 3-Methylcholanthrene and other aryl hydrocarbon receptor agonists directly activate estrogen receptor alpha. Cancer Res. 66, 2459–2467. PubMed

Ahmed S., Valen E., Sandelin A., Matthews J. (2009). Dioxin increases the interaction between aryl hydrocarbon receptor and estrogen receptor alpha at human promoters. Toxicol. Sci. 111, 254–266. PubMed PMC

Ahmed S., Wang A., Celius T., Matthews J. (2014). Zinc finger nuclease-mediated knockout of AHR or ARNT in human breast cancer cells abolishes basal and ligand-dependent regulation of CYP1B1 and differentially affects estrogen receptor alpha transactivation. Toxicol. Sci. 138, 89–103. PubMed

Arcaro K. F., O'Keefe P. W., Yang Y., Clayton W., Gierthy J. F. (1999). Antiestrogenicity of environmental polycyclic aromatic hydrocarbons in human breast cancer cells. Toxicology 133, 115–127. PubMed

Baird W. M., Hooven L. A., Mahadevan B. (2005). Carcinogenic polycyclic aromatic hydrocarbon-DNA adducts and mechanism of action. Environ. Mol. Mutagen. 45, 106–114. PubMed

Beischlag T. V., Perdew G. H. (2005). ER alpha-AHR-ARNT protein-protein interactions mediate estradiol-dependent transrepression of dioxin-inducible gene transcription. J. Biol. Chem. 280, 21607–21611. PubMed

Booc F., Thornton C., Lister A., MacLatchy D., Willett K. L. (2014). Benzo[a]pyrene effects on reproductive endpoints in Fundulus heteroclitus. Toxicol. Sci. 140, 73–82. PubMed PMC

Boström C. E., Gerde P., Hanberg A., Jernström B., Johansson C., Kyrklund T., Rannug A., Törnqvist M., Victorin K., Westerholm R. (2002). Cancer risk assessment, indicators, and guidelines for polycyclic aromatic hydrocarbons in the ambient air. Environ. Health Perspect. 110, 451–488. PubMed PMC

Boylan J. F., Sharp D. M., Leffet L., Bowers A., Pan W. (1999). Analysis of site-specific phosphorylation of the retinoblastoma protein during cell cycle progression. Exp. Cell Res. 248, 110–114. PubMed

Chaloupka K., Krishnan V., Safe S. (1992). Polynuclear aromatic hydrocarbon carcinogens as antiestrogens in MCF-7 human breast cancer cells: role of the Ah receptor. Carcinogenesis 13, 2233–2239. PubMed

Charles G. D., Bartels M. J., Zacharewski T. R., Gollapudi B. B., Freshour N. L., Carney E. W. (2000). Activity of benzo[a]pyrene and its hydroxylated metabolites in an estrogen receptor-alpha reporter gene assay. Toxicol. Sci. 55, 320–326. PubMed

Diamante G., do Amaral E. S. M. G., Menjivar-Cervantes N., Xu E. G., Volz D. C., Dias Bainy A. C., Schlenk D. (2017). Developmental toxicity of hydroxylated chrysene metabolites in zebrafish embryos. Aquat. Toxicol. 189, 77–86. PubMed

Fertuck K. C., Kumar S., Sikka H. C., Matthews J. B., Zacharewski T. R. (2001). Interaction of PAH-related compounds with the alpha and beta isoforms of the estrogen receptor. Toxicol. Lett. 121, 167–177. PubMed

Fertuck K. C., Matthews J. B., Zacharewski T. R. (2001). Hydroxylated benzo[a]pyrene metabolites are responsible for in vitro estrogen receptor-mediated gene expression induced by benzo[a]pyrene, but do not elicit uterotrophic effects in vivo. Toxicol. Sci. 59, 231–240. PubMed

Gozgit J. M., Nestor K. M., Fasco M. J., Pentecost B. T., Arcaro K. F. (2004). Differential action of polycyclic aromatic hydrocarbons on endogenous estrogen-responsive genes and on a transfected estrogen-responsive reporter in MCF-7 cells. Toxicol. Appl. Pharmacol. 196, 58–67. PubMed

Hall J. M., McDonnell D. P. (1999). The estrogen receptor beta-isoform (ERbeta) of the human estrogen receptor modulates ERalpha transcriptional activity and is a key regulator of the cellular response to estrogens and antiestrogens. Endocrinology 140, 5566–5578. PubMed

Hardonnière K., Huc L., Sergent O., Holme J. A., Lagadic-Gossmann D. (2017). Environmental carcinogenesis and pH homeostasis: not only a matter of dysregulated metabolism. Semin. Cancer Biol. 43, 49–65. PubMed

Helle J., Bader M. I., Keiler A. M., Zierau O., Vollmer G., Chittur S. V., Tenniswood M., Kretzschmar G. (2016). Cross-talk in the female rat mammary gland: influence of aryl hydrocarbon receptor on estrogen receptor signaling. Environ. Health Perspect. 124, 601–610. PubMed PMC

Huang B., Butler R., Miao Y., Dai Y., Wu W., Su W., Fujii-Kuriyama Y., Warner M., Gustafsson J. A. (2016). Dysregulation of notch and ERalpha signaling in AhR-/- male mice. Proc. Natl. Acad. Sci. U. S. A. 113, 11883–11888. PubMed PMC

Huang R., Sakamuru S., Martin M. T., Reif D. M., Judson R. S., Houck K. A., Casey W., Hsieh J. H., Shockley K. R., Ceger P., et al. (2015). Profiling of the Tox21 10K compound library for agonists and antagonists of the estrogen receptor alpha signaling pathway. Sci. Rep. 4, 5664. PubMed PMC

Kabátková M., Zapletal O., Tylichová Z., Neča J., Machala M., Milcová A., Topinka J., Kozubík A., Vondráček J. (2015). Inhibition of beta-catenin signalling promotes DNA damage elicited by benzo[a]pyrene in a model of human colon cancer cells via CYP1 deregulation. Mutagenesis 30, 565–576. PubMed

Kamiya M., Toriba A., Onoda Y., Kizu R., Hayakawa K. (2005). Evaluation of estrogenic activities of hydroxylated polycyclic aromatic hydrocarbons in cigarette smoke condensate. Food Chem. Toxicol. 43, 1017–1027. PubMed

Kummer V., Mašková J., Zralý Z., Neča J., Šimečková P., Vondráček J., Machala M. (2008). Estrogenic activity of environmental polycyclic aromatic hydrocarbons in uterus of immature Wistar rats. Toxicol. Lett. 180, 212–221. PubMed

Kuo L. C., Cheng L. C., Lin C. J., Li L. A. (2013). Dioxin and estrogen signaling in lung adenocarcinoma cells with different aryl hydrocarbon receptor/estrogen receptor alpha phenotypes. Am J Respir. Cell Mol. Biol. 49, 1064–1073. PubMed PMC

Labrecque M. P., Takhar M. K., Hollingshead B. D., Prefontaine G. G., Perdew G. H., Beischlag T. V. (2012). Distinct roles for aryl hydrocarbon receptor nuclear translocator and Ah receptor in estrogen-mediated signaling in human cancer cell lines. PLoS One 7, e29545.. PubMed PMC

Lam M. M., Engwall M., Denison M. S., Larsson M. (2018). Methylated polycyclic aromatic hydrocarbons and/or their metabolites are important contributors to the overall estrogenic activity of polycyclic aromatic hydrocarbon-contaminated soils. Environ. Toxicol. Chem. 37, 385–397. PubMed

Legler J., van den Brink C. E., Brouwer A., Murk A. J., van der Saag P. T., Vethaak A. D., van der Burg B. (1999). Development of a stably transfected estrogen receptor-mediated luciferase reporter gene assay in the human T47D breast cancer cell line. Toxicol. Sci. 48, 55–66. PubMed

Liu S., Abdelrahim M., Khan S., Ariazi E., Jordan V. C., Safe S. (2006). Aryl hydrocarbon receptor agonists directly activate estrogen receptor alpha in MCF-7 breast cancer cells. Biol. Chem. 387, 1209–1213. PubMed

Lübcke-von Varel U., Machala M., Ciganek M., Neca J., Pencikova K., Palkova L., Vondracek J., Löffler I., Streck G., Reifferscheid G., et al. (2011). Polar compounds dominate in vitro effects of sediment extracts. Environ. Sci. Technol. 45, 2384–2390. PubMed

Machala M., Vondráček J., Bláha L., Ciganek M., Neča J. (2001). Aryl hydrocarbon receptor-mediated activity of mutagenic polycyclic aromatic hydrocarbons determined using in vitro reporter gene assay. Mutat. Res. 497, 49–62. PubMed

Matthews J., Gustafsson J. A. (2006). Estrogen receptor and aryl hydrocarbon receptor signaling pathways. Nucl. Recept. Signal. 4, e016.. PubMed PMC

Miller M. M., Alyea R. A., LeSommer C., Doheny D. L., Rowley S. M., Childs K. M., Balbuena P., Ross S. M., Dong J., Sun B., et al. (2016). Development of an in vitro assay measuring uterine-specific estrogenic responses for use in chemical safety assessment. Toxicol. Sci. 154, 162–173. PubMed PMC

Moore M., Wang X., Lu Y. F., Wormke M., Craig A., Gerlach J. H., Burghardt R., Barhoumi R., Safe S. (1994). Benzo[a]pyrene-resistant MCF-7 human breast cancer cells. A unique aryl hydrocarbon-nonresponsive clone. J. Biol. Chem. 269, 9. 11751. PubMed

Nebert D. W., Dalton T. P. (2006). The role of cytochrome P450 enzymes in endogenous signalling pathways and environmental carcinogenesis. Nat. Rev. Cancer 6, 947–960. PubMed

Plíšková M., Vondráček J., Canton R. F., Nera J., Kočan A., Petrík J., Trnovec T., Sanderson T., van den Berg M., Machala M. (2005a). Impact of polychlorinated biphenyls contamination on estrogenic activity in human male serum. Environ. Health Perspect. 113, 1277–1284. PubMed PMC

Plíšková M., Vondráček J., Vojtěšek B., Kozubík A., Machala M. (2005b). Deregulation of cell proliferation by polycyclic aromatic hydrocarbons in human breast carcinoma MCF-7 cells reflects both genotoxic and nongenotoxic events. Toxicol. Sci. 83, 246–256. PubMed

Rodgers K. M., Udesky J. O., Rudel R. A., Brody J. G. (2018). Environmental chemicals and breast cancer: an updated review of epidemiological literature informed by biological mechanisms. Environ. Res. 160, 152–182. PubMed

Ruegg J., Swedenborg E., Wahlstrom D., Escande A., Balaguer P., Pettersson K., Pongratz I. (2008). The transcription factor aryl hydrocarbon receptor nuclear translocator functions as an estrogen receptor beta-selective coactivator, and its recruitment to alternative pathways mediates antiestrogenic effects of dioxin. Mol. Endocrinol. 22, 304–316. PubMed PMC

Safe S., Wormke M. (2003). Inhibitory aryl hydrocarbon receptor-estrogen receptor alpha cross-talk and mechanisms of action. Chem. Res. Toxicol. 16, 807–816. PubMed

Shanle E. K., Xu W. (2011). Endocrine disrupting chemicals targeting estrogen receptor signaling: identification and mechanisms of action. Chem. Res. Toxicol. 24, 6–19. PubMed PMC

Shipley J. M., Waxman D. J. (2006). Aryl hydrocarbon receptor-independent activation of estrogen receptor-dependent transcription by 3-methylcholanthrene. Toxicol. Appl. Pharmacol. 213, 87–97. PubMed

Sievers C. K., Shanle E. K., Bradfield C. A., Xu W. (2013). Differential action of monohydroxylated polycyclic aromatic hydrocarbons with estrogen receptors alpha and beta. Toxicol. Sci. 132, 359–367. PubMed PMC

Soto A. M., Sonnenschein C., Chung K. L., Fernandez M. F., Olea N., Serrano F. O. (1995). The E-SCREEN assay as a tool to identify estrogens: an update on estrogenic environmental pollutants. Environ. Health Perspect. 103, 113–122. PubMed PMC

Swedenborg E., Ruegg J., Hillenweck A., Rehnmark S., Faulds M. H., Zalko D., Pongratz I., Pettersson K. (2008). 3-Methylcholanthrene displays dual effects on estrogen receptor (ER) alpha and ER beta signaling in a cell-type specific fashion. Mol. Pharmacol. 73, 575–586. PubMed

Tsai K. S., Yang R. S., Liu S. H. (2004). Benzo[a]pyrene regulates osteoblast proliferation through an estrogen receptor-related cyclooxygenase-2 pathway. Chem. Res. Toxicol. 17, 679–684. PubMed

Van de Wiele T., Vanhaecke L., Boeckaert C., Peru K., Headley J., Verstraete W., Siciliano S. (2004). Human colon microbiota transform polycyclic aromatic hydrocarbons to estrogenic metabolites. Environ. Health Perspect. 113, 6–10. PubMed PMC

van Lipzig M. M., Vermeulen N. P., Gusinu R., Legler J., Frank H., Seidel A., Meerman J. H. (2005). Formation of estrogenic metabolites of benzo[a]pyrene and chrysene by cytochrome P450 activity and their combined and supra-maximal estrogenic activity. Environ. Toxicol. Pharmacol. 19, 41–55. PubMed

Vanparys C., Maras M., Lenjou M., Robbens J., Van Bockstaele D., Blust R., De Coen W. (2006). Flow cytometric cell cycle analysis allows for rapid screening of estrogenicity in MCF-7 breast cancer cells. Toxicol. In Vitro 20, 1238–1248. PubMed

Vondráček J., Kozubík A., Machala M. (2002). Modulation of estrogen receptor-dependent reporter construct activation and G0/G1-S-phase transition by polycyclic aromatic hydrocarbons in human breast carcinoma MCF-7 cells. Toxicol. Sci. 70, 193–201. PubMed

Vondráček J., Machala M., Minksová K., Bláha L., Murk A.J., Kozubík A., Hofmanová J., Hilscherová K., Ulrich R., Ciganek M., et al. (2001). Monitoring river sediments contaminated predominantly with polyaromatic hydrocarbons by chemical and in vitro bioassay techniques. Environ. Toxicol. Chem. 20, 1499–1506. PubMed

Vondráček J., Pěnčíková K., Neča J., Ciganek M., Grycová A., Dvořák Z., Machala M. (2017). Assessment of the aryl hydrocarbon receptor-mediated activities of polycyclic aromatic hydrocarbons in a human cell-based reporter gene assay. Environ. Pollut. 220, 307–316. PubMed

Wihlen B., Ahmed S., Inzunza J., Matthews J. (2009). Estrogen receptor subtype- and promoter-specific modulation of aryl hydrocarbon receptor-dependent transcription. Mol. Cancer Res. 7, 977–986. PubMed

Xue W., Warshawsky D. (2005). Metabolic activation of polycyclic and heterocyclic aromatic hydrocarbons and DNA damage: a review. Toxicol. Appl. Pharmacol. 206, 73–93. PubMed

Zhang Y., Dong S., Wang H., Tao S., Kiyama R. (2016). Biological impact of environmental polycyclic aromatic hydrocarbons (ePAHs) as endocrine disruptors. Environ. Pollut. 213, 809–824. PubMed