Research December 2008 | Volume 116 | Issue 12
Retracted: G-Protein–Coupled Receptor 30 and Estrogen Receptor-α Are Involved in the Proliferative Effects Induced by Atrazine in Ovarian Cancer Cells
Atrazine, one of the most common pesticide contaminants, has been shown to up-regulate aromatase activity in certain estrogen-sensitive tumors without binding or activating the estrogen receptor (ER). Recent investigations have demonstrated that the orphan G-protein–coupled receptor 30 (GPR30), which is structurally unrelated to the ER, mediates rapid actions of 17β-estradiol and environmental estrogens.
Given the ability of atrazine to exert estrogen-like activity in cancer cells, we evaluated the potential of atrazine to signal through GPR30 in stimulating biological responses in cancer cells.
Methods and results:
Atrazine did not transactivate the endogenous ERα in different cancer cell contexts or chimeric proteins encoding the ERα and ERβ hormone-binding domain in gene reporter assays. Moreover, atrazine neither regulated the expression of ERα nor stimulated aromatase activity. Interestingly, atrazine induced extracellular signal-regulated kinase (ERK) phosphorylation and the expression of estrogen target genes. Using specific signaling inhibitors and gene silencing, we demonstrated that atrazine stimulated the proliferation of ovarian cancer cells through the GPR30–epidermal growth factor receptor transduction pathway and the involvement of ERα.
Our results indicate a novel mechanism through which atrazine may exert relevant biological effects in cancer cells. On the basis of the present data, atrazine should be included among the environmental contaminants potentially able to signal via GPR30 in eliciting estrogenic action.
Citation: Albanito L, Lappano R, Madeo A, Chimento A, Prossnitz ER, Cappello AR, Dolce V, Abonante S, Pezzi V, Maggiolini M. 2008. G-Protein–Coupled Receptor 30 and Estrogen Receptor-α Are Involved in the Proliferative Effects Induced by Atrazine in Ovarian Cancer Cells – RETRACTED. Environ Health Perspect 116:1648–1655; http://dx.doi.org/10.1289/ehp.11297
Address correspondence to M. Maggiolini, Department of Pharmaco-Biology, University of Calabria, 87030 Rende (CS), Italy. Telephone: 390984493076. Fax: 390984493458. E-mail: email@example.com
*These authors contributed equally to this work.
This research was supported by grants from the Associazione Italiana per la Ricerca sul Cancro, Ministero dell’Università e Ricerca Scientifica e Tecnologica, and Regione Calabria.
The authors declare they have no competing financial interests.
Received: 28 January 2008
Accepted: 18 July 2008
Advance Publication: 22 July 2008
This paper has been retracted. Click the following link for more information: http://dx.doi.org/10.1289/ehp.11297RET
Albanito L, Madeo A, Lappano R, Vivacqua A, Rago V, Carpino A, et al. 2007. G protein-coupled receptor 30 (GPR30) mediates gene expression changes and growth response to 17beta-estradiol and selective GPR30 ligand G-1 in ovarian cancer cells. Cancer Res 67:1859–1866.
Albanito L, Sisci D, Aquila S, Brunelli E, Vivacqua A, Madeo A, et al. 2008. Epidermal growth factor induces G protein-coupled receptor expression in estrogen receptor-negative breast cancer cells. Endocrinology 149:3799–3808.
Connor K, Howell J, Chen I, Liu H, Berhane K, Sciarretta C, et al. 1996. Failure of chloro-S-triazine-derived compounds to induce estrogen receptor-mediated responses in vivo and in vitro. Fundam Appl Toxicol 30:93–101.
Crain D, Guillette LJ, Rooney AA, Pickford D. 1997. Alterations in steroidogenesis in alligators (Alligator mississippiensis) exposed naturally and experimentally to environmental contaminants. Environ Health Perspect 105:528–533.
Fan W, Yanase T, Morinaga H, Gondo S, Okabe T, Nomura M, et al. 2007a. Atrazine-induced aromatase expression is SF-1 dependent: implications for endocrine disruption in wildlife and reproductive cancers in humans. Environ Health Perspect 115:720–727.
Fan W, Yanase T, Morinaga H, Gondo S, Okabe T, Nomura M, et al. 2007b. Herbicide atrazine activates SF-1 by direct affinity and concomitant co-activators recruitments to induce aromatase expression via promoter II. Biochem Biophys Res Commun 355:1012–1018.
Filardo EJ, Graeber CT, Quinn JA, Resnick MB, Giri D, DeLellis RA, et al. 2006. Distribution of GPR30, a seven membrane-spanning estrogen receptor, in primary breast cancer and its association with clinicopathologic determinants of tumor progression. Clin Cancer Res 12:6359–6366.
Filardo EJ, Quinn JA, Bland KI, Frackelton AR. 2000. Estrogen-induced activation of Erk-1 and Erk-2 requires the G protein-coupled receptor homolog, GPR30, and occurs via trans-activation of the epidermal growth factor receptor through release of HB-EGF. Mol Endocrinol 14:1649–1660.
Filardo EJ, Quinn JA, Frackelton AR, Bland KI. 2002. Estrogen action via the G protein-coupled receptor, GPR30: stimulation of adenylyl cyclase and cAMP-mediated attenuation of the epidermal growth factor receptor-to-MAPK signaling axis. Mol Endocrinol 16:70–84.
Filardo EJ, Quinn JA, Pang C, Graeber CT, Shaw S, Dong J, et al. 2007. Activation of the novel estrogen receptor G protein coupled receptor 30 (GPR30) at the plasma membrane. Endocrinology 148:3236–3245.
Geisinger KR, Kute TE, Pettenati MJ, Welander CE, Dennard Y, Collins LA, et al. 1989. Characterization of a human ovarian carcinoma cell line with estrogen and progesterone receptors. Cancer 63:280–288.
Kniewald J, Mildner P, Kniewald Z. 1979. Effects of s-triazine herbicides on 5α-dihydrotestosterone receptor complex formation, 5α-reductase and 3α-hydroxysteroid dehydrogenase activity at the anterior pituitary level. J Steroid Biochem 11:833–838.
Kniewald J, Mildner P, Kniewald Z. Genazzani E; DiCarlo F; Mainwaring WIP 1980. Effects of s-triazine herbicides on 5α-dihydrotestosterone receptor complex formation in the hypothalamus and ventral prostate. Pharmacological Modulation of Steroid Action New York:Raven Press. 159–169.
Kolpin D, Barbash J, Gilliom R. 1998. Occurence of pesticides in shallow groundwater of the United States: initial results from the National Water-Quality Assessment Program. Environ Sci Technol 32:558–566.
Lehmann TP, Biernacka-Lukanty JM, Saraco N, Langlois D, Li JY, Trzeciak WH. 2005. Temporal pattern of the induction of SF-1 gene expression by the signal transduction pathway involving 3′,5′-cyclic adenosine monophosphate. Acta Biochim Pol 52:485–491.
Loomis AK, Thomas P. 2000. Effects of estrogens and xeno-estrogens on androgen production by Atlantic croaker testes in vitro: evidence for a nongenomic action mediated by an estrogen membrane receptor. Biol Reprod 62:995–1004.
Maggiolini M, Vivacqua A, Fasanella G, Recchia AG, Sisci D, Pezzi V, et al. 2004. The G protein-coupled receptor GPR30 mediates c-fos up-regulation by 17-beta estradiol and phytoestrogens in breast cancer cells. J Biol Chem 279:27008–27016.
McElroy JA, Gangnon RE, Newcomb PA, Kanarek MS, Anderson HA, Brook JV, et al. 2007. Risk of breast cancer for women living in rural areas from adult exposure to atrazine from well water in Wisconsin. J Expo Sci Environ Epidemiol 17:207–214.
Morinaga H, Yanase T, Nomura M, Okabe T, Goto K, Harada N, et al. 2004. A benzimidazole fungicide, benomyl, and its metabolite, carbendazim, induce aromatase activity in a human ovarian granulose-like tumor cell line (KGN). Endocrinology 145:1860–1869.
Müller S, Berg M, Ulrich M, Schwarzenbach RP. 1997. Atrazine and its primary metabolites in Swiss lakes: input characteristics and long-term behavior in the water column. Environ Sci Technol 31:2104–2113.
Nadal A, Ropero AB, Laribi O, Maillet M, Fuentes E, Soria B. 2000. Nongenomic actions of estrogens and xenoestrogens by binding at a plasma membrane receptor unrelated to estrogen receptor alpha and estrogen receptor beta. Proc Natl Acad Sci USA 97:11603–11608.
National Center for Biotechnology Information. 2008. Searching GenBank. Available: http://www.ncbi.nlm.nih.gov/Genbank/GenbankSearch.html [accessed 21 October 2008].
Ruehlmann DO, Steinert JR, Valverde MA, Jacob R, Mann GE. 1988. Environmental estrogenic pollutants induce acute vascular relaxation by inhibiting C-type Ca2+ channels in smooth muscle cells. FASEB J 12:613–619.
Rusiecki JA, De Roos A, Lee WJ, Dosemeci M, Lubin JH, Hoppin JA, et al. 2004. Cancer incidence among pesticide applicators exposed to atrazine in the Agricultural Health Study. J Natl Cancer Inst 96:1375–1382.
Safei R, Katano K, Larson BJ, Samimi G, Holzer AK, Naerdemann W, et al. 2005. Intracellular localization and trafficking of fluorescein-labeled cisplatin in human ovarian carcinoma cells. Clin Cancer Res 11:756–767.
Sanderson JT, Boerma J, Lansbergen G, 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.
Sanderson JT, Letcher RJ, Heneweer M, Giesy JP, Van den Berg M. 2001. Effects of chloro-s-triazine herbicides and metabolites on aromatase activity in various human cell lines and on vitellogenin production in male carp hepatocytes. Environ Health Perspect 109:1027–1031.
Sanderson JT, Seinen W, Giesy JP, van den Berg M. 2000. 2-Chloro-triazine herbicides induce aromatase (CYP19) activity in H295R human adrenocortical carcinoma cells: a novel mechanism for estrogenicity?. Toxicol Sci 54:121–127.
Santagati S, Gianazza E, Agrati P, Vegeto E, Patrone C, Pollio G, et al. 1997. Oligonucleotide squelching reveals the mechanism of estrogen receptor autologous down-regulation. Mol Endocrinol 11:938–949.
Simic B, Kniewald Z, Davies J, Kniewald J. 1991. Reversibility of inhibitory effect of atrazine and lindane on 5-dihydro-testosterone receptor complex formation in rat prostate. Bull Environ Contam Toxicol 46:92–99.
Singleton DW, Feng Y, Burd CJ, Khan SA. 2003. Nongenomic activity and subsequent c-fos induction by estrogen receptor ligands are not sufficient to promote deoxyribonucleic acid synthesis in human endometrial adeno-carcinoma cells. Endocrinology 144:121–128.
Spano L, Tyler C, van Aerle R, Devos P, Mandiki S, Silvestre F, et al. 2004. Effects of atrazine on sex steroid dynamics, plasma vitellogenin concentration and gonad development in adult goldfish (Carassius auratus). Aquat Toxicol 66:369–379.
Stoker TE, Laws S, Guidici D, Cooper R. 2000. The effect of atrazine on puberty in male Wistar rats: an evaluation in the protocol for the assessment of pubertal development and thyroid function. Toxicol Sci 58:50–59.
Stoker TE, Robinette CL, Cooper RL. 1999. Maternal exposure to atrazine during lactation suppresses suckling-induced pro-lactin release and results in prostatitis in the adult offspring. Toxicol Sci 52:68–79.
Tennant MK, Hill DS, Eldridge JC, Wetzel LT, Breckenridge CB, Stevens JT. 1994a. Chloro-s-triazine antagonism of estrogen action: limited interaction with estrogen receptor binding. J Toxicol Environ Health 43:197–211.
Thomas P, Dong J. 2006. Binding and activation of the seven-transmembrane estrogen receptor GPR30 by environmental estrogens: a potential novel mechanism of endocrine disruption. J Steroid Biochem Mol Biol 102:175–179.
Vivacqua A, Bonofiglio D, Albanito L, Madeo A, Rago V, Carpino A, et al. 2006a. 17-beta-Estradiol, genistein, and 4-hydroxytamoxifen induce the proliferation of thyroid cancer cells through the G protein coupled-receptor GPR30. Mol Pharmacol 70:1414–1423.
Vivacqua A, Bonofiglio D, Recchia AG, Musti AM, Picard D, Andò S, et al. 2006b. The G protein-coupled receptor GPR30 mediates the proliferative effects induced by 17beta-estradiol and hydroxytamoxifen in endometrial cancer cells. Mol Endocrinol 20:631–646.
Wetzel LT, Luempert LG, Breckenridge CB, Tisdel MO, Stevens JT, Thakur AK, et al. 1994. Chronic effects of atrazine on estrus and mammary tumor formation in female Sprague-Dawley and Fischer 344 rats. J Toxicol Environ Health 43:169–182.
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