Skip to main content

Advertisement

SpringerLink
Log in

Effects of single pesticides and binary pesticide mixtures on estrone production in H295R cells

  • In vitro systems
  • Published:
Archives of Toxicology Aims and scope Submit manuscript

Abstract

The aim of the present study was to determine whether the human adrenocortical carcinoma cell line H295R can be used as an in vitro test system to investigate the effects of binary pesticide combinations on estrone production as biological endpoint. In the first step ten pesticides selected according to a tiered approach were tested individually. The anilinopyrimidines cyprodinil and pyrimethanil as well as the dicarboximides iprodione and procymidone increased estrone concentration, while the triazoles myclobutanil and tebuconazole as well as the strobilurins azoxystrobin and kresoxim-methyl decreased estrone concentration in the supernatant of H295R cells. The N-methylcarbamate methomyl did not show any effects, and the phthalimide captan reduced estrone concentration unspecifically due to its detrimental impact on cellular viability. When cyprodinil and pyrimethanil, which belong to the same chemical group and increase estrone production, were combined, in most of the cases the overall effect was solely determined by the most potent compound in the mixture (i.e., cyprodinil). When cyprodinil and procymidone, which belong to different chemical groups but increase estrone production, were combined, in most cases an additive effect was observed. When cyprodinil, which increased estrone production, was combined with either myclobutanil or azoxystrobin, which decreased estrone production, the overall effect of the mixture was in most cases either entirely determined by myclobutanil or at least partially modulated by azoxystrobin. In conclusion, H295R cells appear to be an adequate in vitro test system to study the effect of combining two pesticides affecting estrone production.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8

Similar content being viewed by others

References

  • Allen JA, Shankara T, Janus P, Buck S, Diemer T, Hales KH, Hales DB (2006) Energized, polarized, and actively respiring mitochondria are required for acute Leydig cell steroidogenesis. Endocrinology 147:3924–3935

    Article  CAS  PubMed  Google Scholar 

  • Andersen HR, Vinggard AM, Rasmussen TH, Gjermandsen IM, Bonefeld-Jørgensen EVA (2002) Effects of currently used pesticides in assays for estrogenicity, androgenicity, and aromatase activity in vitro. Toxicol Appl Pharmacol 179:1–12

    Article  CAS  PubMed  Google Scholar 

  • Andersen HR, Bonefeld Jørgensen EC, Nielsen F, Jarfeldt K, Jayatissa MN, Vinggaard AM (2006) Estrogenic effects in vitro and in vivo of the fungicide fenarimol. Toxicol Lett 163:142–152

    Article  CAS  PubMed  Google Scholar 

  • Becker WF, von Jagow G, Anke T, Steglich W (1981) Oudemansin, strobilurin A, strobilurin B and myxothiazol: new inhibitors of the bc1 segment of the respiratory chain with an E-β-methoxyacrylate system as common structural element. FEBS Lett 132:329–333

    Article  CAS  PubMed  Google Scholar 

  • Bonefeld Jørgensen EC, Autrup H, Hansen JC (1997) Effect of toxaphene on estrogen receptor functions in human breast cancer cells. Carcinogenesis 18:1651–1654

    Article  PubMed  Google Scholar 

  • Boobis AR, Ossendorp BC, Banasiak U, Hamey PY, Sebestyen I, Moretto A (2008) Cumulative risk assessment of pesticide residues in food. Toxicol Lett 180:137–150

    Article  CAS  PubMed  Google Scholar 

  • Boon PE, van Klaveren JD (2003) Cumulative exposure to acetylcholinesterase inhibiting compounds in the Dutch population and young children. Toxicity equivalence approach with acephate and phosmet as index compounds. RIKILT, Wageningen. Available on-line at http://www.rikilt.wur.nl/NR/rdonlyres/BDEEDD31-F58C-47EB-A0AA-23CB9956CE18/18704/R2003003.pdf

  • Budimir M, Plesnicar M, Kljajić R (1976) Effects of the herbicide 2,4-DB and fungicide captan on reactions of mitochondria and chloroplasts. Arch Environ Contam Toxicol 4:166–174

    Article  CAS  PubMed  Google Scholar 

  • BVL (Federal Office of Consumer Protection and Food Safety, Germany) (2012a) Archiv der Berichte zu Pflanzenschutzmittelrückständen. Available on-line at: http://www.bvl.bund.de/DE/01_Lebensmittel/01_Aufgaben/02_AmtlicheLebensmittelueberwachung/09_PSMRueckstaende/01_nb_psm/nbpsm_gesamtuebersicht/nbpsm_gesamtuebersicht_berichte.html;jsessionid=B6B71D9A8E8EDC3E3386847815F4DE2B.1_cid094?nn=1399970

  • BVL (Federal Office of Consumer Protection and Food Safety, Germany) (2012b) Nationale Berichterstattung Pflanzenschutzmittelrückstände in Lebensmitteln 2010. Available on-line at: http://www.bvl.bund.de/DE/01_Lebensmittel/01_Aufgaben/02_AmtlicheLebensmittelueberwachung/09_PSMRueckstaende/01_nb_psm/nbpsm_2010/lm_nbpsm_bericht_2010_basepage.html?nn=1399970

  • Commission Regulation (EC) No 2076/2002 of 20 November 2002 extending the time period referred to in Article 8(2) of Council Directive 91/414/EEC and concerning the non-inclusion of certain active substances in Annex I to that Directive and the withdrawal of authorisations for plant protection products containing these substances (OJ L 319, 23.11.2002)

  • COT (UK Committee on Toxicity of chemicals in food, consumer products and the environment) (2002) Risk assessment of mixtures of pesticides and similar substances (WIGRAMP). Available on-line at: http://cot.food.gov.uk/pdfs/reportindexed.pdf

  • Council Directive 79/117/EEC of 21 December 1978 prohibiting the placing on the market and use of plant protection products containing certain active substances (OJ L 33, 08.02.1979), now repealed by Regulation (EC) No 1107/2009 of the European Parliament and of the Council of 21 October 2009 concerning the placing of plant protection products on the market and repealing Council Directives 79/117/EEC and 91/414/EEC (OJ L 309, 24.11.2009)

  • CVUA (Chemisches und Veterinäruntersuchungsamt Stuttgart, Germany) (2007) Toxikologische Bewertung von Mehrfachrückständen in Obst und Gemüse. Available on-line at: http://www.cvuas.de/pub/beitrag.asp?Thema_ID=5&ID=648&subid=1&Pdf=True

  • EFSA (European Food Safety Authority) (2008) Scientific opinion of the panel on plant protection products and their residues (PPR Panel) on a request from the EFSA evaluate the suitability of existing methodologies and, if appropriate, the identification of new approaches to assess cumulative and synergistic risks from pesticides to human health with a view to set MRLs for those pesticides in the frame of Regulation (EC) 396/2005. EFSA J 704:1–84. Available on-line at: http://www.efsa.europa.eu/en/efsajournal/doc/705.pdf

  • EFSA (European Food Safety Authority) (2009) Scientific Opinion on risk assessment for a selected group of pesticides from the triazole group to test possible methodologies to assess cumulative effects from exposure through food from these pesticides on human health. EFSA J 7:1167. Available on-line at: http://www.efsa.europa.eu/en/efsajournal/doc/1167.pdf

  • EPA (US Environmental Protection Agency) (2002) Guidance on Cumulative risk assessment of pesticide chemicals that have a common mechanism of toxicity. Washington, DC. Available on-line at: http://www.epa.gov/oppfead1/trac/science/cumulative_guidance.pdf

  • EPA (US Environmental Protection Agency) (2006a) Acetochlor/Alachlor: Cumulative risk assessment for the chloroacetanilides. Washington, DC. Available on-line at: http://www.epa.gov/pesticides/cumulative/chloro_cumulative_risk.pdf

  • EPA (US Environmental Protection Agency) (2006b) Triazine cumulative risk assessment. Washington, DC. Available on-line at: http://www.epa.gov/oppsrrd1/ REDs/triazine_cumulative_risk.pdf

  • EPA (US Environmental Protection Agency) (2006c) Organophosphorus cumulative risk assessment—2006 Update. Washington, DC. Available on-line at: http://www.epa.gov/oppsrrd1/cumulative/2006-op/op_cra_main.pdf

  • EPA (US Environmental Protection Agency) (2007) Revised N-methyl carbamate cumulative risk assessment. Washington, DC. Available on-line at: http://www.epa.gov/oppsrrd1/REDs/nmc_revised_cra.pdf

  • Gazdar AF, Oie HK, Shackleton CH, Chen TR, Triche TJ, Myers CE, Chrousos GP, Brennan MF, Stein CA, La Rocca RV (1990) Establishment and characterization of a human adrenocortical carcinoma cell line that expresses multiple pathways of steroid biosynthesis. Cancer Res 50:5488–5496

    CAS  PubMed  Google Scholar 

  • Goetz AK, Rockett JC, Ren HZ, Thillainadarajah I, Dix DJ (2009) Inhibition of rat and human steroidogenesis by triazole antifungals. Syst Biol Reprod Med 55:214–226

    Article  CAS  PubMed  Google Scholar 

  • ILSI (The International Life Sciences Institute) (1999) A framework for cumulative risk assessment. ILSI Press, Washington

    Google Scholar 

  • Jensen AF, Petersen A, Granby K (2003) Cumulative risk assessment of the intake of organophosphorus and carbamate pesticides in the Danish diet. Food Addit Contam 20:776–785

    Article  CAS  PubMed  Google Scholar 

  • Kjærstad MB, Taxvig C, Nellemann C, Vinggaard AM, Andersen HR (2010a) Endocrine disrupting effects in vitro of conazole antifungals used as pesticides and pharmaceuticals. Reprod Toxicol 30:573–582

    Article  PubMed  Google Scholar 

  • Kjærstad MB, Taxvig C, Andersen HR, Nellemann C (2010b) Mixture effects of endocrine disrupting compounds in vitro. Int J Androl 33:425–433

    Article  PubMed  Google Scholar 

  • Lu SY, Liao JW, Kuo ML, Wang SC, Hwang JS, Ueng TH (2004) Endocrine-disrupting activity in carbendazim-induced reproductive and developmental toxicity in rats. J Toxicol Environ Health A 67:1501–1515

    Article  CAS  PubMed  Google Scholar 

  • Mason JI, Murry BA, Olcott M, Sheets JJ (1985) Imidazole antimycotics: inhibitors of steroid aromatase. Biochem Pharmacol 34:1087–1092

    Article  CAS  PubMed  Google Scholar 

  • Midzak AS, Chen H, Aon MA, Papadopoulos V, Zirkin BR (2011) ATP synthesis, mitochondrial function, and steroid biosynthesis in rodent primary and tumor Leydig cells. Biol Reprod 84:976–985

    Article  CAS  PubMed  Google Scholar 

  • Montanaro D, Maggiolini M, Recchia AG, Sirianni R, Aquila S, Barzon L, Fallo F, Ando S, Pezzi V (2005) Antiestrogens upregulate estrogen receptor β expression and inhibit adrenocortical H295R cell proliferation. J Mol Endocrinol 35:245–256

    Article  CAS  PubMed  Google Scholar 

  • Ostby J, Kelce WR, Lambright C, Wolf CJ, Mann P, Gray LE Jr (1999) The fungicide procymidone alters sexual differentiation in the male rat by acting as an androgen-receptor antagonist in vivo and in vitro. Toxicol Ind Health 15:80–93

    CAS  PubMed  Google Scholar 

  • Payne AH, Hales DB (2004) Overview of steroidogenic enzymes in the pathway from cholesterol to active steroid hormones. Endocr Rev 25:947–970

    Article  CAS  PubMed  Google Scholar 

  • Pfeil R (2007) Fungizide. In: Dunkelberg H, Gebel T, Hartwig A (eds) Handbuch der lebensmitteltoxikologie, vol 3. Wiley-VCH Verlag, Weinheim, pp 1357–1410

    Google Scholar 

  • Radice S, Fumagalli R, Chiesara E, Ferraris M, Frigerio S, Marabini L (2004) Estrogenic activity of procymidone in rainbow trout (Oncorhynchus mykiss) hepatocytes: a possible mechanism of action. Chem Biol Interact 147:185–193

    Article  CAS  PubMed  Google Scholar 

  • Radice S, Chiesara E, Frigerio S, Fumagalli R, Parolaro D, Rubino T, Marabini L (2006) Estrogenic effect of procymidone through activation of MAPK in MCF-7 breast carcinoma cell line. Life Sci 78:2716–2723

    Article  CAS  PubMed  Google Scholar 

  • Regulation (EC) No. 396/2005 of the European Parliament and of the Council of 23 February 2005 on maximum residue levels of pesticides in or on food and feed of plant and animal origin and amending Council Directive 91/414/EEC. OJ L70/1, 16.3.2005

  • Rone MB, Fan J, Papadopoulos V (2009) Cholesterol transport in steroid biosynthesis: role of protein–protein interactions and implications in disease states. Biochim Biophys Acta 1791:646–658

    Article  CAS  PubMed  Google Scholar 

  • Sanderson JT, Boerma J, Lansbergen GW, van den Berg M (2002) Induction and inhibition of aromatase (CYP19) activity by various classes of pesticides in H295R human adrenocortical carcinoma cells. Toxicol Appl Pharmacol 182:44–54

    Article  CAS  PubMed  Google Scholar 

  • Somjen D, Stern N, Knoll E, Sharon O, Gayer B, Kulik T, Kohen F (2003) Carboxy derivatives of isoflavones as affinity carriers for cytotoxic drug targeting in adrenocortical H295R carcinoma cells. J Endocrinol 179:395–403

    Article  CAS  PubMed  Google Scholar 

  • Sonneveld E, Jansen HJ, Riteco JAC, Brouwer A, van der Burg B (2005) Development of androgen- and estrogen-responsive bioassays, members of a panel of human cell line-based highly selective steroid responsive bioassays. Toxicol Sci 83:136–148

    Article  CAS  PubMed  Google Scholar 

  • Suzuki T, Nojiri H, Isono H, Ochi T (2004) Oxidative damages in isolated rat hepatocytes treated with the organochlorine fungicides captan, dichlofluanid and chlorothalonil. Toxicology 204:97–107

    Article  CAS  PubMed  Google Scholar 

  • Taxvig C, Hass U, Axelstad M, Dalgaard M, Boberg J, Andersen HR, Vinggaard AM (2007) Endocrine-disrupting activities in vivo of the fungicides tebuconazole and epoxiconazole. Toxicol Sci 100:464–473

    Article  CAS  PubMed  Google Scholar 

  • van der Burg B, Winter R, Man H, Vangenechten C, Weimer M, Berckmans P, Witters H, van der Linden S (2010a) Optimization and prevalidation of the in vitro AR CALUX method to test androgenic and antiandrogenic activity of compounds. Reprod Toxicol 30:18–24

    Article  PubMed  Google Scholar 

  • van der Burg B, Winter R, Weimer M, Berckmans P, Suzuki G, Gijsbers L, Jonas A, van der Linden S, Witters H, Aarts J, Legler J, Kopp-Schneider A, Bremer S (2010b) Optimization and prevalidation of the in vitro ERα CALUX method to test estrogenic and antiestrogenic activity of compounds. Reprod Toxicol 30:73–80

    Article  PubMed  Google Scholar 

  • Villeneuve DL, Ankley GT, Makynen EA, Blake LS, Greene KJ, Higley B, Newsted JL, Giesy JP, Hecker M (2007) Comparison of fathead minnow ovary explant and H295R cell-based steroidogenesis assays for identifying endocrine-active chemicals. Ecotoxicol Environ Saf 68:20–32

    Article  CAS  PubMed  Google Scholar 

  • Vinggaard AM, Jørgensen EC, Larsen JC (1999a) Rapid and sensitive reporter gene assays for detection of antiandrogenic and estrogenic effects of environmental chemicals. Toxicol Appl Pharmacol 155:150–160

    Article  CAS  PubMed  Google Scholar 

  • Vinggaard AM, Breinholt V, Larsen JC (1999b) Screening of selected pesticides for oestrogen receptor activation in vitro. Food Addit Contam 16:533–542

    Article  CAS  PubMed  Google Scholar 

  • Vinggaard AM, Hnida C, Breinholt V, Larsen JC (2000) Screening of selected pesticides for inhibition of CYP19 aromatase activity in vitro. Toxicol In Vitro 14:227–234

    Article  CAS  PubMed  Google Scholar 

  • Vinggaard AM, Jacobsen H, Metzdorff SB, Andersen HR, Nellemann C (2005) Antiandrogenic effects in short-term in vivo studies of the fungicide fenarimol. Toxicology 207:21–34

    Article  CAS  PubMed  Google Scholar 

  • VKM (Norwegian Scientific Committee for Food Safety) (2008) Combined toxic effects of multiple chemical exposures. Report 1. Available on-line at: http://vkm.no/dav/15ec300082.pdf

Download references

Acknowledgments

We thank Dr. Iris Suckrau (LAVES, Oldenburg, Germany) for providing the data on residue contents of fruits and vegetables.

Conflict of interest

The authors state that they have no conflict of interest.

Author information

Author notes

  1. Wiebke Prutner

    Present address: Federal Institute for Occupational Safety and Health (BAuA), Friedrich-Henkel-Weg 1-25, 44149, Dortmund, Germany

  2. Gerd Hamscher

    Present address: Institute of Food Chemistry and Food Biotechnology, Justus-Liebig-University, Heinrich-Buff-Ring 58, 35392, Giessen, Germany

Authors and Affiliations

  1. Institute for Food Toxicology and Analytical Chemistry, University of Veterinary Medicine Hannover, Bischofsholer Damm 15, 30173, Hannover, Germany

    Wiebke Prutner, Petra Nicken, Gerd Hamscher & Pablo Steinberg

  2. Lower Saxony State Office for Consumer Protection and Food Safety (LAVES), Röverskamp 5, 26203, Wardenburg, Germany

    Eberhard Haunhorst

Authors
  1. Wiebke Prutner
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Petra Nicken
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eberhard Haunhorst
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gerd Hamscher
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Pablo Steinberg
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Pablo Steinberg.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Prutner, W., Nicken, P., Haunhorst, E. et al. Effects of single pesticides and binary pesticide mixtures on estrone production in H295R cells. Arch Toxicol 87, 2201–2214 (2013). https://doi.org/10.1007/s00204-013-1081-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00204-013-1081-x

Keywords

Search

Navigation