Steroid hormones bind to highly specific nuclear receptors, regulating gene expression that results in normal fetal growth and development and/or in normal adult physiological function. Many industrial and agricultural chemicals may bind one or more nuclear receptors, acting as mimics of steroid hormones, and are called endocrine disruptive chemicals (EDC) because they alter the expression of endocrine-regulated genes. A widely used fungicide, Enable (fenbuconazole), was evaluated to examine its capacity to alter endocrine-regulated gene expression. Cells of an oestrogen-dependent human breast cancer-derived line, MCF-7, were treated with a range, 0.033-3.3 ppb (ng/mL), of Enable, and gene expression was compared to that of untreated cells. Microarray analysis using a chip with 600 gene spots showed downregulation of eight genes and upregulation of 34 genes in cells treated with 3.3 ppb of Enable, compared to untreated cells. Specific genes were selected for consideration. Real-time PCR confirmed results obtained from analysis of the microarray data for the genes phenol sulphotransferase (PST), intercellular adhesion molecule-1 (ICAM-1), transforming growth factor b-3 (TGF b-3) and calreticulin. These studies were designed to provide base-line data on the gene expression-altering capacity of a specific chemical at a low dose, and will allow assessment of the possible deleterious effects that may be caused in human cells by exposure to the agricultural chemical Enable.
Berthois, Y., Katzenellenbogen, J. A. and Katzenellenbogen, B. S.1986: Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proceedings of the National Academy of Sciences of the United States of America83, 2496-2500.
2.
Brix, L. A., Nicoll, R., Zhu, X.et al.1998: Structural and functional characterisation of human sulfotransferases. Chemico-Biological Interactions109(1-3), 123-127.
3.
Burns, K., Duggan, B., Atkinson, A.et al.1994: Modulation of gene expression by calreticulin binding to the glucocorticoid receptor. Nature367, 476-480.
4.
Caulin-Glaser, T., Watson, C. A., Pardi, R.et al.1996: Effects of 17beta-estradiol on cytokine-induced endothelial cell adhesion molecule expression. The Journal of Clinical Investigation98, 36-42.
5.
Constable, J. D. and Hatch, M. C.1985: Reproductive effects of herbicide exposure in Vietnam: recent studies by the Vietnamese and others. Teratogenesis, Carcinogenesis and Mutagenesis5, 231-250.
6.
Coughtrie, M. W., Sharp, S., Maxwell, K.et al.1998: Biology and function of the reversible sulphonation pathway catalyzed by human sulfotransferases and sulfatases. Chemico-biological Interactions109, 3-27.
7.
Crofton, K. M., Kodavanti, P. R. S., Derr-Yellin, E. C.et al.2000: PCBs, thyroid hormones, and toxicity in rats: cross-fostering experiments demonstrate the impact of postnatal lactation exposure. Toxicological Sciences57, 131-140.
8.
Davidson, Lupton, J. R., Mishowsky, E.et al.2003: Quantification of human intestinal gene expression profiles using exfoliated colonocytes: a pilot study. Biomarkers8, 51-61.
9.
Dong, X., Pulido, J. S., Ou, T.et al.2003: Differentiation of human neural stem cells into retinal cells. Neuroreport14, 143-146.
10.
Dunker, N. and Krieglstein, K.2002: Tgfbeta2-/- Tgfbeta3//- double knockout mice display severe midline fusion defects and early embryonic lethality. Anatomy and Embryology206, 73-83.
Fuller, P.1991: The steroid receptor superfamily: mechanisms of diversity. The FASEB Journal5, 3092-3099.
14.
Gilbert, M. E., Mundy, W. R. and Crofton, K. W.2000: Spatial learning and long-term potentiation in the dentate gyrus of the hippocampus in animals developmentally exposed to Aroclor 1254. Toxicological Sciences57, 102-111.
15.
Glatt, H., Boeing, H., Engelke, C. E.et al.2001: Human cytosolic sulphotransferases: genetics, characteristics, toxicological aspects. Mutation Research482, 27-40.
16.
Glover, V., Littlewood, J., Sondler, M.et al.1983: Biochemical predisposition to dietary migraine: the role of phenolsulphotransferase. Headache23, 53-58.
17.
Jackson, T. F. and Halbert, F. L.1974: A toxic syndrome associated with the feeding of polybrominated biphenyl-contaminated protein concentrate to dairy cattle. Journal of the American Veterinary Medical Association165, 437-439.
18.
Jakowlew, S. B., Zakowicz, H. and Moody, T. W.2000: Retinoic acid down-regulates VPAC(1) receptors and TGF-beta 3 but up-regulates TGF-beta 2 in lung cancer cells. Peptides21, 1831-1837.
19.
Kashimoto, T. and Miyata, H.1987: Differences between Yusho and other kinds of poisoning involving only PCBs. In Waid, J. S., editor, PCBs and the environment, Boca Raton, FL: CRC Press.
20.
Lawrence, C., Reilly, A., Quyickenton, P., Greenwald, P., Page, W. and Kuntz, A.1985: Mortality patterns of New York State Vietnam veterans. American Journal of Public Health75, 277-277.
21.
Lieberman, B. A.1997: The estrogen receptor activity cycle: dependence on multiple protein-protein interaction. Crit. Rec. In Endocr. Gene Expr.7, 43-59.
22.
Lui, W. Y., Lee, W. M. and Cheng, C. Y.2003: TGF-betas: their role in testicular function and Sertoli cell tight junction dynamics. International Journal of Andrology26, 147-160.
23.
Lynch, J. and Michalak, M.2003: Calreticulin is an upstream regulator of calcineurin. Biochemical and Biophysical Research Communications311, 1173-1179.
24.
Mery, L., Mesaeli, N., Michalak, M.et al.1996: Over expression of calreticulin increases intracellular Ca2storage and decreases store-operated Ca2 influx. The Journal of Biological Chemistry271, 9332-9339.
25.
Michalak, M., Robert Parker, J. M. and Opas, M.2002: Ca2signaling and calcium binding chaperones of the endoplasmic reticulum. Cell Calcium32, 269-278.
26.
Nguyen, T. O., Capra, J. D. and Sontheimer, R. D.1996: Calreticulin is transcriptionally upregulated by heat shock, calcium and heavy metals. Molecular Immunology33, 379-386.
27.
O’Malley, B. and Tsai, M-J.1992: Molecular pathways of steroid receptor action. Biology of Reproduction46, 163-167.
28.
Ogawa, Y., Hirakawak, K., Nokata, B.et al.1998: Expression of intercellular adhesion molecule-1 in invasive breast cancer reflects low growth potential, negative lymph node involvement, and good prognosis. Clinical Cancer Research4, 31-36.
29.
Poland, A. and Knutson, J. C.1982: Polychlorinated biphenyls (PCBs) and polybrominated biphenyls (PBBs): biochemistry, toxicology and mechanisms of activity. Annual Review of Pharmacology and Toxicology22, 517-517.
30.
Roegge, C. S., Seo, B.-W., Crofton, K. M. and Schantz, S. L.2000: Gestational-lactational exposure to Aroclor 1254 impairs radial-arm maze performance in male rats. Toxicological Sciences57, 121-130.
31.
Runge-Morris, M.1998: Regulation of sulfotransferase gene expression by glucocorticoid hormones and xenobiotics in primary rat hepatocyte culture. Chemico-biological Interactions109, 315-327.
32.
Safe, S.1990: 2,3,7,8-Tetrachlorodibenzo-p-dioxin, and related halogenated aromatic hydrocarbons: examination of mechanisms of toxicity. Toxicology21, 51-88.
33.
Santostefano, M. J., Wang, X., Richardson, V. M., Ross, D. G., DeVito, M. J. and Birnbaum, L. S.1998: A pharmacodynamic analysis of TCDD-induced cytochrome P450 gene expression in multiple tissues: dose-and time-dependent effects. Toxicology and Applied Pharmacology151, 294-310.
34.
Seegal, R. F.2000: The neurotoxicological consequences of developmental exposure to PCBs. Toxicological Sciences57, 1-3.
35.
Seth, P., Lunetta, K. L., Bell, D. W.et al.2000: Phenol sulfotransferases: hormonal regulation, polymorphism, and age of onset of breast cancer. Cancer Research60, 6859-6863.
36.
Stanciu, L. A. and Djukanovic, R.1998: The role of ICAM-1 on T-cells in the pathogenesis of asthma. The European Respiratory Journal11, 949-957.
37.
St-Arnaud, R., Prud’homme, J., Leung-Hagerteijn, C.et al.1995: Constitutive expression of calreticulin in osteoblasts inhibits mineralization. The Journal of Cell Biology131, 1351-1359.
38.
Tamura, H.-O., Taniguchi, K., Hayashi, Y.et al.2001: Expression profiling of sulfotransferases in human cell lines derived from extra-hepatic tissues. Biological and Pharmaceutical Bulletin24, 1258-1258.
39.
Truss, M. and Beato, M.1993: Steroid hormone receptors: interaction with deoxyribonucleic acid and transcription factors. Endocrine Reviews14(4), 450-479.
40.
Van de Stolpe, A. and Van der Saag, P. T.1996: Intercellular adhesion molecule-1. Journal of Molecular Medicine74, 13-33.
41.
Wang, Y., Wang, L. and Wang, S.1998: Study on expression of transforming growth factor-beta 3 in the liver tissues. Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi12, 61-63.
42.
Waxman, D. A.1999: P450 gene induction by structurally diverse xenochemicals: central role of nuclear receptors CAR, PXR, and PPAR. Archives of Biochemistry and Biophysics369, 11-29.
43.
Whitlock, J. P. Jr.1986: The regulation of cytochrome P450 gene expression. Annual Review of Pharmacology and Toxicology26, 333-333.
44.
Wood, A.2003: Compendium of pesticide common names. Copyright # 1995-2003 Alan Wood. Retrieved 9 July, 2003, from: http://www.hclrss.demon.co.uk
45.
Yusuf-Makagiansar, H., Anderson, M. E., Yakovleva, T. V.et al.2002: Inhibition of LFA-1/ICAM-1 and VLA-4/VCAM-1 as a therapeutic approach to inflammation and autoimmune diseases. Medicinal Research Reviews22, 146-167.
46.
Zarn, J. A., Bruschweiler, B. J. and Schlatter, J. R.2003: Azole fungicides affect mammalian steroidogenesis by inhibiting sterol 14 alpha-demethylase and aromatase. Environmental Health Perspectives111, 255-261.
47.
Zhu, N. and Wang, Z.1999: Calreticulin expression is associated with androgen regulation of the sensitivity to calcium ionophore-induced apoptosis in LNCaP prostate cancer cells. Cancer Research59, 1896-1902.
