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AroER tri-screen™ is a novel functional assay to estimate both estrogenic and estrogen precursor activity of chemicals or biological specimens

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Abstract

The purpose of the study is to define AroER tri-screen’s utility for identifying endocrine-disrupting chemicals (EDCs) that target aromatase and/or estrogen receptor (ER), and to measure the total estrogenic activity in biological specimens. ER-positive, aromatase-expressing MCF-7 breast cancer cells were stably transfected with an estrogen responsive element (ERE)-driven luciferase reporter plasmid to yield a new high-throughput screening platform—the AroER tri-screen. AroER tri-screen was capable of identifying estrogen precursors, such as cortodoxone, which function as estrogens through a two-step conversion process in aromatase-expressing tissue. Furthermore, the system proved useful for assessing EDC activity in biologically relevant samples. Estimating these activities is critical because natural estrogens and estrogenic EDCs are important factors in ER-positive breast cancer risk. As our research demonstrates, incorporating functionally active aromatase into the AroER tri-screen produces a powerful and unique tool to (1) identify new EDCs targeting aromatase and/or ER; (2) discover novel EDCs activated by aromatase; and (3) estimate overall estrogenic activities in biological samples as a potential intermediate risk factor for breast cancer.

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References

  1. Diamanti-Kandarakis E, Bourguignon JP, Giudice LC, Hauser R, Prins GS, Soto AM, Zoeller RT, Gore AC (2009) Endocrine-disrupting chemicals: an Endocrine Society scientific statement. Endocr Rev 30(4):293–342. doi:10.1210/er.2009-0002

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  2. Casals-Casas C, Desvergne B (2011) Endocrine disruptors: from endocrine to metabolic disruption. Annu Rev Physiol 73:135–162. doi:10.1146/annurev-physiol-012110-142200

    Article  CAS  PubMed  Google Scholar 

  3. Chen S, Zhou D, Hsin LY, Kanaya N, Wong C, Yip R, Sakamuru S, Xia M, Yuan YC, Witt K, Teng C (2014) AroER Tri-screen is a biologically relevant assay for endocrine disrupting chemicals modulating the activity of aromatase and/or the estrogen receptor. Toxicol Sci. doi:10.1093/toxsci/kfu023

    Google Scholar 

  4. Tox-21 10K library (2012) http://www.epa.gov/ncct/dsstox/sdf_tox21s.html

  5. Rotroff DM, Dix DJ, Houck KA, Knudsen TB, Martin MT, McLaurin KW, Reif DM, Crofton KM, Singh AV, Xia M, Huang R, Judson RS (2013) Using in vitro high throughput screening assays to identify potential endocrine-disrupting chemicals. Environ Health Perspect 121(1):7–14. doi:10.1289/ehp.1205065

    PubMed Central  PubMed  Google Scholar 

  6. Hjelmborg PS, Ghisari M, Bonefeld-Jorgensen EC (2006) SPE-HPLC purification of endocrine-disrupting compounds from human serum for assessment of xenoestrogenic activity. Anal Bioanal Chem 385(5):875–887. doi:10.1007/s00216-006-0463-9

    Article  CAS  PubMed  Google Scholar 

  7. Kruger T, Spano M, Long M, Eleuteri P, Rescia M, Hjelmborg PS, Manicardi GC, Bizzaro D, Giwercman A, Toft G, Bonde JP, Bonefeld-Jorgensen EC (2008) Xenobiotic activity in serum and sperm chromatin integrity in European and inuit populations. Mol Reprod Dev 75(4):669–680. doi:10.1002/mrd.20747

    Article  CAS  PubMed  Google Scholar 

  8. Kanno Y, Okada H, Kobayashi T, Takenaka T, Suzuki H (2007) Effects of endocrine disrupting substance on estrogen receptor gene transcription in dialysis patients. Ther Apher Dial 11(4):262–265. doi:10.1111/j.1744-9987.2007.00472.x

    Article  CAS  PubMed  Google Scholar 

  9. Lim VW, Li J, Gong Y, Yuan JM, Wu TS, Hammond GL, Jin A, Koh WP, Yong EL (2012) Serum free estradiol and estrogen receptor-alpha mediated activity are related to decreased incident hip fractures in older women. Bone 50(6):1311–1316. doi:10.1016/j.bone.2012.03.006

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Widschwendter M, Lichtenberg-Frate H, Hasenbrink G, Schwarzer S, Dawnay A, Lam A, Menon U, Apostolidou S, Raum E, Stegmaier C, Jacobs IJ, Brenner H (2009) Serum oestrogen receptor alpha and beta bioactivity are independently associated with breast cancer: a proof of principle study. Br J Cancer 101(1):160–165. doi:10.1038/sj.bjc.6605106

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. Fourkala EO, Zaikin A, Burnell M, Gentry-Maharaj A, Ford J, Gunu R, Soromani C, Hasenbrink G, Jacobs I, Dawnay A, Widschwendter M, Lichtenberg-Frate H, Menon U (2012) Association of serum sex steroid receptor bioactivity and sex steroid hormones with breast cancer risk in postmenopausal women. Endocr Relat Cancer 19(2):137–147. doi:10.1530/ERC-11-0310

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  12. Lim VW, Li J, Gong Y, Jin A, Yuan JM, Yong EL, Koh WP (2014) Serum estrogen receptor bioactivity and breast cancer risk among postmenopausal women. Endocr Relat Cancer 21(2):263–273. doi:10.1530/ERC-13-0233

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  13. Shen J, Xu L, Fang H, Richard AM, Bray JD, Judson RS, Zhou G, Colatsky TJ, Aungst JL, Teng C, Harris SC, Ge W, Dai SY, Su Z, Jacobs AC, Harrouk W, Perkins R, Tong W, Hong H (2013) EADB: an estrogenic activity database for assessing potential endocrine activity. Toxicol Sci 135(2):277–291. doi:10.1093/toxsci/kft164

    Article  CAS  PubMed  Google Scholar 

  14. Auzeby A, Bogdan A, Touitou Y (1991) Evidence for a new biologic pathway of androstenedione synthesis from 11-deoxycortisol. Steroids 56(1):33–36

    Article  CAS  PubMed  Google Scholar 

  15. Key T, Appleby P, Barnes I, Reeves G, Endogenous H, Breast Cancer Collaborative G (2002) Endogenous sex hormones and breast cancer in postmenopausal women: reanalysis of nine prospective studies. J Natl Cancer Inst 94(8):606–616

    Article  CAS  PubMed  Google Scholar 

  16. Kelly CM, Juurlink DN, Gomes T, Duong-Hua M, Pritchard KI, Austin PC, Paszat LF (2010) Selective serotonin reuptake inhibitors and breast cancer mortality in women receiving tamoxifen: a population based cohort study. BMJ 340:c693. doi:10.1136/bmj.c693

    Article  PubMed Central  PubMed  Google Scholar 

  17. To SQ, Knower KC, Cheung V, Simpson ER, Clyne CD (2014) Transcriptional control of local estrogen formation by aromatase in the breast. J Steroid Biochem Mol Biol. doi:10.1016/j.jsbmb.2014.05.004

    PubMed  Google Scholar 

  18. Sperry TS, Thomas P (1999) Characterization of two nuclear androgen receptors in Atlantic croaker: comparison of their biochemical properties and binding specificities. Endocrinology 140(4):1602–1611. doi:10.1210/endo.140.4.6631

    CAS  PubMed  Google Scholar 

  19. Piferrer F, Baker IJ, Donaldson EM (1993) Effects of natural, synthetic, aromatizable, and nonaromatizable androgens in inducing male sex differentiation in genotypic female chinook salmon (Oncorhynchus tshawytscha). Gen Comp Endocrinol 91(1):59–65. doi:10.1006/gcen.1993.1104

    Article  CAS  PubMed  Google Scholar 

  20. Verheul HA, Schot LP, Deckers GH, Schuurs AH (1986) Effects of tibolone, lynestrenol, ethylestrenol, and desogestrel on autoimmune disorders in NZB/W mice. Clin Immunol Immunopathol 38(2):198–208

    Article  CAS  PubMed  Google Scholar 

  21. Allenby F, Jeyasingh K, Calnan J (1973) Ethyloestrenol and postoperative venous thrombosis. Lancet 2(7819):38–39

    Article  CAS  PubMed  Google Scholar 

  22. Van Puymbroeck M, Kuilman ME, Maas RF, Witkamp RF, Leyssens L, Van Miert AS, Hendriks L, Vanderzande D, Adriaensens P, Jacobs MP, Raus J (1999) 17alpha-ethyl-5beta-estrane-3alpha, 17beta-diol, a biological marker for the abuse of norethandrolone and ethylestrenol in slaughter cattle. J Chromatogr B Biomed Sci Appl 728(2):217–232

    Article  PubMed  Google Scholar 

  23. Dey P, Barros RP, Warner M, Strom A, Gustafsson JA (2013) Insight into the mechanisms of action of estrogen receptor beta in the breast, prostate, colon, and CNS. J Mol Endocrinol 51(3):T61–T74. doi:10.1530/JME-13-0150

    Article  CAS  PubMed  Google Scholar 

  24. Cappon GD, Cook JC, Hurtt ME (2003) Relationship between cyclooxygenase 1 and 2 selective inhibitors and fetal development when administered to rats and rabbits during the sensitive periods for heart development and midline closure. Birth Defects Res B 68(1):47–56. doi:10.1002/bdrb.10008

    Article  CAS  Google Scholar 

  25. Maitra S, Baidya DK, Khanna P, Ray BR, Panda SS, Bajpai M (2014) Acute perioperative pain in neonates: an evidence-based review of neurophysiology and management. Acta Anaesthesiol Taiwanica 52(1):30–37. doi:10.1016/j.aat.2014.02.004

    Article  Google Scholar 

  26. Ward RM, Kearns GL (2013) Proton pump inhibitors in pediatrics : mechanism of action, pharmacokinetics, pharmacogenetics, and pharmacodynamics. Paediatr Drugs 15(2):119–131. doi:10.1007/s40272-013-0012-x

    Article  PubMed Central  PubMed  Google Scholar 

  27. Katchy A, Pinto C, Jonsson P, Nguyen-Vu T, Pandelova M, Riu A, Schramm KW, Samarov D, Gustafsson JA, Bondesson M, Williams C (2014) Coexposure to phytoestrogens and bisphenol a mimics estrogenic effects in an additive manner. Toxicol Sci 138(1):21–35. doi:10.1093/toxsci/kft271

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  28. Wong SP, Li J, Shen P, Gong Y, Yap SP, Yong EL (2007) Ultrasensitive cell-based bioassay for the measurement of global estrogenic activity of flavonoid mixtures revealing additive, restrictive, and enhanced actions in binary and higher order combinations. Assay Drug Dev Technol 5(3):355–362. doi:10.1089/adt.2007.056

    Article  CAS  PubMed  Google Scholar 

  29. Kaaks R, Rinaldi S, Key TJ, Berrino F, Peeters PH, Biessy C, Dossus L, Lukanova A, Bingham S, Khaw KT, Allen NE, Bueno-de-Mesquita HB, van Gils CH, Grobbee D, Boeing H, Lahmann PH, Nagel G, Chang-Claude J, Clavel-Chapelon F, Fournier A, Thiebaut A, Gonzalez CA, Quiros JR, Tormo MJ, Ardanaz E, Amiano P, Krogh V, Palli D, Panico S, Tumino R, Vineis P, Trichopoulou A, Kalapothaki V, Trichopoulos D, Ferrari P, Norat T, Saracci R, Riboli E (2005) Postmenopausal serum androgens, oestrogens and breast cancer risk: the European prospective investigation into cancer and nutrition. Endocr Relat Cancer 12(4):1071–1082. doi:10.1677/erc.1.01038

    Article  CAS  PubMed  Google Scholar 

  30. Ziegler RG, Faupel-Badger JM, Sue LY, Fuhrman BJ, Falk RT, Boyd-Morin J, Henderson MK, Hoover RN, Veenstra TD, Keefer LK, Xu X (2010) A new approach to measuring estrogen exposure and metabolism in epidemiologic studies. J Steroid Biochem Mol Biol 121(3–5):538–545. doi:10.1016/j.jsbmb.2010.03.068

    Article  CAS  PubMed  Google Scholar 

  31. Endogenous H, Breast Cancer Collaborative G, Key TJ, Appleby PN, Reeves GK, Travis RC, Alberg AJ, Barricarte A, Berrino F, Krogh V, Sieri S, Brinton LA, Dorgan JF, Dossus L, Dowsett M, Eliassen AH, Fortner RT, Hankinson SE, Helzlsouer KJ, Hoffman-Bolton J, Comstock GW, Kaaks R, Kahle LL, Muti P, Overvad K, Peeters PH, Riboli E, Rinaldi S, Rollison DE, Stanczyk FZ, Trichopoulos D, Tworoger SS, Vineis P (2013) Sex hormones and risk of breast cancer in premenopausal women: a collaborative reanalysis of individual participant data from seven prospective studies. Lancet Oncol 14(10):1009–1019. doi:10.1016/S1470-2045(13)70301-2

    Article  Google Scholar 

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Acknowledgments

The authors would like to thank Talisman, Ian, PhD and Kelly Yeo for assistance in editing the manuscript and luciferase assay, respectively. We also would like to thank Susan Markel in the the Analytical Pharmacology Core for her performance of liquid chromatography and mass spectrometry analysis. This work was supported by California Breast Cancer Research Program [Grant number 17UB-8701] and the National Cancer Institute (P30 CA033572), NIH/NIEHS (ESO8258). In addition, we would like to thank Dr. Joanne Mortimer for her support and comments.

Conflict of interest

The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Department of Cancer Biology, Beckman Research Institute of the City of Hope, 1500 East Duarte Road, Duarte, CA, 91010, USA

    Noriko Kanaya, Duc M. Nguyen, Hannah Lu, Yuan-Zhong Wang, Li-Yu Hsin, Tim Synold & Shiuan Chen

  2. Environmental Chemistry Branch, California Department of Toxic Substances Control, Sacramento, CA, 94710, USA

    Myrto Petreas & Weihong Guo

  3. The Cancer Prevention Institute of California, Fremont, CA, 94538, USA

    David Nelson & Peggy Reynolds

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  1. Noriko Kanaya
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Correspondence to Shiuan Chen.

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Kanaya, N., Nguyen, D.M., Lu, H. et al. AroER tri-screen™ is a novel functional assay to estimate both estrogenic and estrogen precursor activity of chemicals or biological specimens. Breast Cancer Res Treat 151, 335–345 (2015). https://doi.org/10.1007/s10549-015-3398-z

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  • DOI: https://doi.org/10.1007/s10549-015-3398-z

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