Glyphosate-based herbicides are toxic and endocrine disruptors in human cell lines

Toxicology

Volumn 262, Issue 3, 2009, Pages 184-191

  Gasnier, Céline ^a   Dumont, Coralie ^b   Benachour, Nora ^a   Clair, Emilie ^a   Chagnon, Marie Christine ^b   Séralini, Gilles Eric ^a  

^a UNIVERSITÉ DE CAEN   (France)

^b UNIVERSITÉ DE BOURGOGNE   (France)

Author keywords

Endocrine disruptor; Glyphosate; HepG2; Roundup; Sexual steroids

Indexed keywords

ANDROGEN; ANDROGEN RECEPTOR; AROMATASE; DNA; ENDOCRINE DISRUPTOR; ESTROGEN; ESTROGEN RECEPTOR ALPHA; ESTROGEN RECEPTOR BETA; GLYPHOSATE; HERBICIDE; MESSENGER RNA;

ARTICLE; CARCINOGENICITY; CELL ASSAY; CELL STRAIN HEPG2; COMET ASSAY; CONCENTRATION (PARAMETERS); CONTROLLED STUDY; CYTOTOXICITY; DNA DAMAGE; DRUG FORMULATION; ENZYME ACTIVITY; FOOD CONTAMINATION; GENOTOXICITY; HUMAN; HUMAN CELL; MUTAGENICITY; PRIORITY JOURNAL; REPORTER GENE; REPRODUCTIVE TOXICITY; TRANSCRIPTION REGULATION;

ADOLESCENT; AROMATASE; CELL LINE, TUMOR; COMET ASSAY; DNA DAMAGE; DOSE-RESPONSE RELATIONSHIP, DRUG; ENDOCRINE DISRUPTORS; ESTROGEN RECEPTOR ALPHA; ESTROGEN RECEPTOR BETA; GENES, REPORTER; GLYCINE; HERBICIDES; HUMANS; MALE; PESTICIDE RESIDUES; RECEPTORS, ANDROGEN; RNA, MESSENGER; TOXICITY TESTS; TRANSCRIPTION, GENETIC;

EID: 67651014408     PISSN: 0300483X     EISSN: None     Source Type: Journal    
DOI: 10.1016/j.tox.2009.06.006     Document Type: Article

References (94)

1. Acquavella J.F., Bruce H., Alexander B.H., Mandel J.S., Gustin C., Baker B., Champan P., and Bleeke M. Glyphosate biomonitoring for farmers and their families: results from the farm family exposure study. Environ. Health Perspect. 112 (2004) 321-326
2. Acquavella J.F., Alexander B.H., Mandel J.S., Burns C.J., and Gustin C. Exposure misclassification in studies of agricultural pesticides. Epidemiology 17 (2006) 69-74
3. Amrhein N., Deus B., Gehrke P., and Steinrüken H.C. The site of the inhibition of the shikimate pathway by glyphosate. Plant Physiol. 66 (1980) 830-834
4. Arnold S.F., Vonier P.M., Collins B.M., Klotz D.M., Guillette Jr. L.J., and McLachlan J.A. In vitro synergistic interaction of alligator and human estrogen receptors with combinations of environmental chemicals. Environ. Health Perspect. 105 (1997) 615-618
5. Bellé R., Le Bouffant R., Morales J., Cosson B., Cormier P., and Mulner-Lorillon O. Sea urchin embryo, DNA-damaged cell cycle checkpoint and the mechanisms initiating cancer development. J. Soc. Biol. 201 (2007) 317-327
6. Benachour N., Moslemi S., Sipahutar H., and Séralini G.E. Cytotoxic effects and aromatase inhibition by xenobiotic endocrine disrupters alone and in combination. Toxicol. Appl. Pharmacol. 222 (2007) 129-140
7. Benachour N., Sipahutar H., Moslemi S., Gasnier C., Travert C., and Séralini G.E. Time- and dose-dependent effects of Roundup on human embryonic and placental cells. Environ. Contam. Toxicol. 53 (2007) 126-133
8. Benachour N., and Séralini G.E. Glyphosate formulations induce apoptosis and necrosis in human umbilical, embryonic, and placental cells. Chem. Res. Toxicol. 22 (2009) 97-105
9. Beuret C.J., Zirulnik F., and Gimenez M.S. Effect of the herbicide glyphosate on liver lipoperoxidation in pregnat rats and their fetuses. Reprod. Toxicol. 19 (2005) 501-504
10. Blum J.L., Nyagode B.A., James M.O., and Denslow N.D. Effects of the pesticide methoxychlor on gene expression in the liver and testes of the male largemouth bass (Micropterus salmoides). Aquat. Toxicol. 86 (2008) 469-1469
11. Borenfreund E., and Puerner J.A. Toxicity determined in vitro by morphological alterations and neutral red absorption. Toxicol. Lett. 24 (1984) 119-124
12. Bourguiba S., Lambard S., and Carreau S. Steroids control the Aromatase gene expression in purified germ cells from the adult male rat. J. Mol. Endocrinol. 31 (2003) 83-94
13. Bradford M.M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72 (1976) 248-254
14. Cabaton N., Dumont C., Severin I., Perdu E., Zalko D., Cherkaoui-Malki M., and Chagnon M.C. Genotoxic and endocrine activities of bis(hydroxyphenyl)methane (bisphenol F) and its derivatives in the HepG2 cell line. Toxicology 255 (2009) 15-24
15. Cadet J., Douki T., Gasparutto D., and Ravanat J.L. Oxidative damage to DNA: formation, measurement and biochemical features. Mutat. Res. 531 (2003) 5-23
16. ® on the fish Prochilodus lineatus. Mutat. Res. 655 (2008) 41-46
17. Cavas T., and Könen S. Detection of cytogenetic and DNA damage in peripheral erythrocytes of goldfish (Carassius auratus) exposed to a glyphosate formulation using the micronucleus test and the comet assay. Mutagenesis 22 (2007) 263-268
18. Clements C., Ralph S., and Petras M. Genotoxicity of select herbicides in Rana catesbeiana tadpoles using the alkaline single-cell gel DNA electrophoresis (comet) assay. Environ. Mol. Mutagen. 29 (1997) 277-288
19. Clive, J. 2009. Global status of biotech/GM Crops: 2008. ISAAA Brief 39.
20. Collins A.R. The comet assay for NA damage and repair. Principles, applications and limitations. Mol. Biotechnol. 26 (2004) 249-261
21. Collins A.R., Oscoz A.A., Brunborg G., Gaivao I., Giovannelli L., Kruszewski M., Smith C.C., and Stetina R. The comet assay: topical issues. Mutagenesis 23 (2008) 143-151
22. Contardo-Jara V., Klingelmann E., and Wiegand C. Bioaccumulation of glyphosate and its formulation Roundup in lumbriculus variegates and its effects on biotransformation and antioxidant enzymes. Environ. Pollut. 157 (2008) 57-63
23. Corbin C.J., Graham-Lorence S., McPhaul M., Mason J.I., Mendelson C.R., and Simpson E.R. Islation of a full-lengh cDNA insert encoding human aromatase system cytochrom P-450 and its expression in nonsteroidogenic cells. Proc. Natl. Acad. Sci. 85 (1988) 8948-8952
24. Cox C. Glyphosate (Roundup). J. Pest. Reform. 18 (1998) 3-17
25. Crouch S.P.M., Kozlowski R., Slater K.J., and Fletcher J. The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity. J. Immunol. Methods 160 (1993) 81-88
26. Dallegrave E., Mantese F.D., Oliveira R.T., Andrade A.J.M., Dalsenter P.R., and Langeloh A. Pre- and postnatal toxicity of the commercial glyphosate formulation in Wistar rats. Reprod. Toxicol. 81 (2007) 665-673
27. Daruich J., Zirulnik F., and Gimenez M.S. Effect of the herbicide glyphosate on enzymatic activity in pregnant rats and their fetuses. Environ. Res. Sect. 85 (2001) 226-231
28. De Roos A., Blair A., Rusiecki J.A., Hoppin J.A., Svec M., Dosemeci M., Sandler D.P., and Alavanja M.C. Cancer incidence among glyphosate-exposed pesticide applicators in the agricultural health study. Environ. Health Pespect. 113 (2005) 49-54
29. Denizot F., and Lang R. Rapid colorimetric assay for cell growth and survival modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J. Immunol. Methods 89 (1986) 271-277
30. Dimitrov B.D., Gadeva P.G., Benova D.K., and Bineva M.V. Comparative genotoxicity of the herbicide roundup, stomp and reglone in plant and mammalian test systems. Mutagenesis 21 (2006) 375-382
31. Dintinger T., Gaillard J.L., Moslemi S., Zwain I., and Silberzhan P. Androgen and 19-norandrogen aromatization by equine and human placental microsomes. J. Steroid Biochem. 33 (1989) 949-954
32. Edwards T.M., and Myers J.P. Environmental exposures and gene regulation in disease etiology. Environ. Health Perspect. 115 (2007) 1264-1270
33. Feron V.J., Cassee F.R., Groten J.P., van Vliet P.W., and van Zorge J.A. International issues on human health effects of exposure to chemical mixtures. Environ. Health Perspect. 110 (2002) 893-899
34. Franz J.E. In: Grossbard E., and Atkinson D. (Eds). The Herbicide Glyphosate (1985), Butterworth, Boston, MA 1-17
35. Hall R.E., Birrell S.N., Tilley W.D., and Sutherland R.L. MDA-MB-453, an androgen-responsive human breast carcinoma cell line with high level androgen receptor expression. Eur. J. Cancer. 30 (1994) 484-490
36. Hall J.M., and McDonnell D.P. The estrogen receptor β-isoform (ERβ) of the human estrogen receptor modulates ERα transcriptional activity and is a key regulator of the cellular response to estrogens and antiestrogens. Endocrinology 140 (1999) 5566-5578
37. Heydens W.F., Healy C.E., Hotz K.J., Kier L.D., Martens M.A., Wilson A.G.E., and Farmer D.N. Genotoxic potential of glyphosate formulations: mode-of-action investigations. J. Agric. Food Chem. 56 (2008) 1517-1523
38. Hodgson E., and Rose R.L. Human metabolic interactions of environmental chemicals. J. Biochem. Mol. Toxicol. 21 (2007) 182-186
39. Hokanson R., Fudge R., Chowdhary R., and Busbee D. Alteration of estrogen-regulated gene expression in human cells induced by the agricultural and horticultural herbicide glyphosate. Hum. Exp. Toxicol. 26 (2007) 747-752
40. IFEN. Les pesticides dans les eaux. Dossier 5 (2006) 15-20
41. Kaya B., Creus A., Yanikoǧlu A., Cabré O., and Marcos R. Use of the Drosophila wing spot test in the genotoxicity testing of different herbicides. Environ. Mol. Mutagen. 36 (2000) 40-46
42. Kirkland D.J., Aardema M., Banduhn N., Carmichael P., Fautz R., Meunier J.R., and Pfuhler S. In vitro approaches to develop weight of evidence (WoE) and mode of action (MoA) discussions with positive in vitro genotoxicity results. Mutagenesis 22 (2007) 161-175
43. Knasmüller S., Mersh-Sundermann V., Kevekordes S., Darroudi F., Huber W.W., Hoelzl C., Bichler J., and Majer B.J. Use of human-derived liver cell lines for the detection of environmental and dietary genotoxicants; current state of knowledge. Toxicology 198 (2004) 315-328
44. 2) and nongenotoxic environmental chemicals in human fibroblasts. Toxicol. Lett. 147 (2004) 35-43
45. Lioi M.B., Scarfì M.R., Santoro A., Barbieri R., Zeni O., Di Berardino D., and Ursini M.V. Genotoxicity and oxidative stress induced by pesticide exposure in bovine lymphocyte cultures in vitro. Mutat. Res. 403 (1998) 13-20
46. Malatesta M., Perdoni F., Santin G., Battistelli S., Muller S., and Biggiogera M. Hepatoma tissue culture (HTC) cells as a model for investigating the effects of low concentrations of herbicide on cell structure and function. Toxicol. In Vitro. 22 (2008) 1853-1860
47. Mañas F., Peralta L., Raviolo J., Garcia Ovando H., Weyers A., Ugnia L., Gonzales Cid M., Larripa I., and Gorla N. Genotoxicity of AMPA, the environmental metabolite of glyphosate, assessed by the Comet assay and cytogenetic tests. Ecotoxicol. Environ. Saf. 72 (2008) 834-837
48. Marc J., Mulner-Lorillon O., Boulben S., Hureau D., Durand G., and Bellé R. Pesticide roundup provokes cell division dysfunction at the level of CDK1/Cyclin B activation. Chem. Res. Toxicol. 15 (2002) 326-331
49. Marc J., Mulner-Lorillon O., and Bellé R. Glyphosate-based pesticides affect cell cycle regulation. Biol. Cell. 96 (2004) 245-249
50. Marc J., Le Breton M., Cormier P., Morales J., Bellé R., and Mulner-Lorillon O. A glyphosate-based pesticide impinges on transcription. Toxicol. Appl. Pharmacol. 203 (2005) 1-8
51. Martin-Skilton R., Saborido-Rey F., and Porte C. Endocrine alteration and other biochemical responses in juvenile turbot exposed to the Prestige fuel oil. Sci. Tot. Environ. 404 (2008) 68-76
52. Mattews J., and Gustafsson J.A. Estrogen receptor and aryl hydrocarbon receptor signaling pathways. Nucl. Recept. Signal. 4 (2006) 1-4
53. Monroy C.M., Cortes A.C., Sicard D.M., and de Restrepo H.G. Cytotoxicity and genotoxicity of human cells exposed in vitro to glyphosate. Biomedica 25 (2005) 335-345
54. Moslemi S., and Séralini G.E. Estrogens and breast cancer: aromatase activity disruption. Trends In Breast Cancer Research (2005) pp. 101-127
55. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65 (1983) 55-63
56. O'Brien J., Wilson I., Orton T., and Pognan F. Investigation of the Alamar Blue (resazurin) fluorescent dye for the assessment of mammalian cell cytotoxicity. Eur. J. Biochem. 267 (2000) 5421-5426
57. Oliveira A.G., Telles L.F., Hess R.A., Mahecha G.A.B., and Oliveira C.A. Effects of herbicide Roundup on the epididymal region of drakes Anas platyrhynchos. Reprod. Toxicol. 23 (2007) 182-191
58. Olmstead A.W., Kosian P.A., Korte J.J., Holcombe G.W., Woodis K.K., and Degitz S.J. Sex reversal of the amphibian, Xenopus tropicalis, following larval exposure to an aromatase inhibitor. Acquat. Toxicol. 91 (2009) 143-150
59. Ozcan-Arican G., and Ozalpan A. Evaluation of the effect of paclitaxel, epirubicin and tamoxifen by cell kinetics parameters in estrogen-receptor-positive ehrlich ascites tumor (eat) cells growing in vitro. Acta Biol. Hung. 58 (2007) 49-59
60. Paris F., Balaguer P., Térouanne B., Servant N., Lacoste C., Cravedi J.P., Nicolas J.C., and Sultan C. Phenylphenols, biphenols, bisphenol-A and 4-ter-octylphenol exhibit α and β estrogen activities and antiandrogen activity in reporter cell lines. Mol. Cell. Endocrinol. 193 (2002) 43-49
61. Peluso M., Munnia A., Bolognesi C., and Parodi S. 32P-postlabeling detection of DNA adducts in mice treated with the herbicide roundup. Environ. Mol. Mutagen. 31 (1998) 55-59
62. Poletta G.L., Larriera A., Kleinsorge E., and Mudry M.D. Caiman latirostris (broad-snouted caiman) as a sentinel organism for genotoxic monitoring: Basal values determination of micronucleus and comet assay. Mutat. Res. 650 (2008) 202-209
63. Relyea R.A. A cocktail of contaminants: how mixtures of pesticides at low concentrations affect aquatic communities. Oecologia 159 (2008) 363-376
64. Richard S., Moslemi S., Sipahutar H., Benachour N., and Séralini G.E. Differential effects of glyphosate and roundup on human placental cells and aromatase. Environ. Health Perspect. 113 (2005) 716-720
65. Rogan W.J., and Ragan N.B. Some evidence of effects of environmental chemicals on the endocrine system in children. Int. J. Hyg. Environ. Health 210 (2007) 659-667
66. Rokutanda N., Iwasaki T., Odawara H., Nagaoka R., Miyazaki W., Takeshita A., Koibuchi Y., Horiguchi J., Shimokawa N., Iino Y., Morishista Y., and Koibuchi N. Augmentation of estrogen receptor-mediated transcription by steroid and xenobiotic receptor. Endocrinology 33 (2008) 305-316
67. Salaberria I., Hansen B.H., Asensio V., Olsvik P.A., Andersen R.A., and Jenssen B.M. Effects of atrazine on hepatic metabolism and endocrine homeostasis in rainbow trout (Oncorhynchus mykiss). Toxicol. Appl. Pharmacol. 234 (2009) 98-106
68. Savitz D., Arbuckle T., Kaczor D., and Curtis K.M. Male pesticide exposure and pregnancy outcome. Am. J. Epidemiol. 146 (1997) 1025-1036
69. Szarek J., Siwicki A., Andrzejewska A., Terech-Majewska E., and Banaszkiewicz T. Effects of the herbicide Roundup on the ultrastructural pattern of hepatocytes in carp (Cyprinus carpio). Mar. Environ. Res. 50 (2000) 263-266
70. Scatena R., Messana I., Martorana G.E., Gozzo M.L., Lippa S., Maccaglia A., Bottoni P., Vincenzoni F., Nocca G., Castagnola M., and Giardina B. Mitochondrial damage and metabolic compensatory mechanisms induced by hyperoxia in the U-937 cell line. J. Biochem. Mol. Biol. 37 (2004) 454-459
71. Séralini G.E., and Moslemi S. Aromatase inhibitors: past, present and future. Mol. Cell. Endocrinol. 178 (2001) 117-131
72. Sheleer C.Q., Dudley M.W., and Khan S.A. Environmental estrogens induce transcriptionally active estrogen receptor dimmers in yeast: activity potentiated by the coactivator RIP140. Environ. Health Perspect. 108 (2000) 97-103
73. Sibley R., Hatoum-Mokdad H., Schoenleber R., Musza L., Stirtan W., Marrero D., Carley W., Xiao H., and Dumas J. A novel estrogen receptor ligand template. Bioorg. Med. Biochem. Lett. 13 (2003) 1919-1922
74. Simard J., Luthy I., Guay J., Bélanger A., and Labrie F. Characteristics of interaction of the antiandrogen flutamide with the androgen receptor in various target tissues. Mol. Cell Endo. 44 (1986) 261-270
75. Simpson E.R., Mahendroo M.S., Means G.D., Kilgore M.W., Hinshelwood M.M., and Grham-Lorenze S. Aromatase Cytochrome P450, the enzyme responsible for estrogen biosynthesis. Endocrionol. Rev. 15 (1994) 342-355
76. Simpson E.R., Clyne C., Rubin G., Boon W.C., Robertson K., Britt K., Speed C., and Jones M. Aromatase a brief overview. Annu. Rev. Physiol. 64 (2002) 93-127
77. Singh N.P., McCoy M.T., Tice R.R., and Schneider E.L. A simple technique for quantition of low levels of DNA damage in individual cells. Exp. Cell Res. 175 (1988) 184-191
78. Squirrell D., and Murphy J. Rapid detection of very low numbers of micro-organisms using adenylate kinase as a cell marker. A Practical Guide to Industrial Uses of ATP Luminescence in Rapid Microbiology (1997), Cara Technology Ltd, Lingfield, Surrey, UK pp. 107-113
79. Sultan C., Balaguer P., Terouanne B., Georget V., Paris F., Jeandel C., Lumbroso S., and Nicolas J.C. Environmental xenoestrogens, antiandrogens and disorders of male sexual differentiation. Mol. Cel. Endocrinol. 178 (2001) 99-105
80. Takahashi M., Horie M., and Aoba N. Analysis of glyphosate and its metabolite, aminomethylphosphonic acid, in agricultural products by HPLC. Shokuhin Eiseigaku Zasshi 42 (2001) 304-308
81. Thompson E.A., and Siiteri P.K. Utilization of oxygen and reduced nicotinamide adenine dinucleotide phosphate by human placental microsomes during aromatization of androstenedione. J. Biol. Chem. 249 (1974) 5364-5372
82. ®) on the toxicity and bioavailability of metals to Ceriodaphnia dubia. Environ. Pollut. 138 (2005) 59-68
83. Tzukerman M.T., Esty A., Santiso-Mere D., Danielian P., Parker M.G., Stein R.B., Pike J.W., and McDonnell D.P. Human estrogen receptor transactivational capacity is determined by both cellular and promoter context and mediated by two functionally distinct intramolecular regions. Mol. Endocrinol. 8 (1994) 21-30
84. Urani C., Doldi M., Crippa S., and Camatini M. Human-derived cell lines to study xenobiotic metabolism. Chemosphere 37 (1998) 2785-2795
85. U.S. Environment Protection Agency, 1998. Endocrine disruptor screening and testing advisory committee (ECSTAC) final report, Washington.
86. Valentin-Severin I., Le Hegarat L., Lebon A.M., Lhuguenot J.C., and Chagnon M.C. Use of HepG2 cell line for direct or indirect mutagens screening: comparative investigation between comet and micronucleus assays. Mutat. Res. 536 (2003) 79-90
87. Walsh L.P., McCormick C., Martin C., and Stocco D.M. Roundup inhibits steroidogenesis by disrupting steroidogenic acute regulatory (StAR) protein expression. Environ. Health Perspect. 108 (2000) 769-776
88. Watson C.S., Bulayeva N.N., Wozniak A.L., and Alyea R.A. Xenoestrogens are potent activators of nongenomic responses. Steroids 72 (2006) 124-134
89. Westerink, and Schoonen. Cytochrome P450 enzyme levels in HepG2 cells and cryopreserved primary human hepatocytes and their induction in HepG2 cells. Toxicol In Vitro. 21 (2007) 1581-1591
90. Williams G.M., Kroes R., and Munro I.C. Safety evaluation and risk of the herbicide Roundup and its active ingredient, glyphosate, for humans. Regul. Toxicol. Pharmacol. 31 (2000) 117-165
91. Wilson V.S., Bobsein K., Lambright C.R., and Gray L.E. A novel cell line, MDA-kb2, that stably expresses an androgen- and glucocorticoid-responsive reporter for the detection of hormone receptor agonists and antagonists. Toxicol. Sci. 66 (2002) 69-81
92. Wilson V.S., Blystone C.R., Hotchkiss A.K., Rider C.V., and Gray Jr. L.E. Diverse mechanisms of anti-androgen action: impact on male rat reproductive tract development. Int. J. Androl. 31 (2008) 178-187
93. Xu L.C., Sun H., Chen J.F., Bian Q., Qian J., Song L., and Wang X.R. Evaluation of androgen receptor transcriptional activities of bisphenol A, octylphenol and nonylphenol in vitro. Toxicology 216 (2005) 197-203
94. Yousef M.I., Salem M.H., Ibrahim H.Z., Helmi S., Seehy M.A., and Bertheussen K. Toxic effects of carbofuran and glyphosate on semen characteristics in rabbits. J. Environ. Sci. Health B 30 (1995) 513-534