Serum harvested from heifers one month post-zeranol implantation stimulates MCF-7 breast cancer cell growth

Ye,Weiping; Xu,Pingping; Zhong,Saiyi; Threlfall,Walter R.; Frasure,Christopher; Feng,Eric; Li,Hong; Lin,Shu-Hong; Liu,Jie-Yu; Lin,Young C.

doi:10.3892/etm.2010.155

Serum harvested from heifers one month post-zeranol implantation stimulates MCF-7 breast cancer cell growth

Authors:
- Weiping Ye
- Pingping Xu
- Saiyi Zhong
- Walter R. Threlfall
- Christopher Frasure
- Eric Feng
- Hong Li
- Shu-Hong Lin
- Jie-Yu Liu
- Young C. Lin
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Affiliations: Laboratory of Reproductive and Molecular Endocrinology, College of Veterinary Medicine, The Ohio State University, Columbus OH 43210, USA, Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus OH 43210, USA, Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus OH 43210, USA
Published online on: September 29, 2010 https://doi.org/10.3892/etm.2010.155
Pages: 963-968

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Abstract

Breast cancer is a serious disease in the US. Numerous risk factors have been linked to this disease. The safety of using growth promoters, such as zeranol, remains under debate due to the lack of sufficient in vitro and in vivo evidence. Using CellTiter 96™ Aqueous Non-Radioactive Cell Proliferation assay, real-time PCR and Western blot analysis, we evaluated the effects of sera harvested from experimental and control heifers before and after one month of zeranol implantation on the growth of human breast cancer cell line MCF-7 as well as the involved mechanisms. We found that sera harvested from the heifers following one month of zeranol implantation were more mitogenically potent in stimulating the proliferation of MCF-7 cells when compared to sera harvested from the same heifers before zeranol implantation and the control heifers. Further investigation found that dextran-coated charcoal suppressed the stimulating effect of the sera on MCF-7 cell growth. The mechanisms involved in the MCF-7 cell proliferation stimulated by zeranol-containing sera may include up-regulation of cyclin D1 and down-regulation of p53 and p21 expression at the mRNA and protein levels in the cells. The results suggest that the consumption of beef products containing biologically active residues of zeranol or its metabolites is a risk linked to breast cancer development. Further investigation is required in order to clarify this critical issue.

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1.	Jemal A, Siegel R, Ward E, et al: Cancer statistics, 2009. CA Cancer J Clin. 59:225–249. 2009. View Article : Google Scholar
2.	Brekelmans CT: Risk factors and risk reduction of breast and ovarian cancer. Curr Opin Obstet Gynecol. 15:63–68. 2003. View Article : Google Scholar : PubMed/NCBI
3.	Delort L, Satih S, Kwiatkowski F, et al: Evaluation of breast cancer risk in a multigenic model including low penetrance genes involved in xenobiotic and estrogen metabolisms. Nutr Cancer. 62:243–251. 2010. View Article : Google Scholar : PubMed/NCBI
4.	Thonney ML: Growth, feed efficiency and variation of individually fed Angus, polled Hereford and Holstein steers. J Anim Sci. 65:1–8. 1987.PubMed/NCBI
5.	Prichard DL, Hargrove DD, Olson TA, et al: Effects of creep feeding, zeranol implants and breed type on beef production: I. Calf and cow performance. J Anim Sci. 67:609–616. 1989.PubMed/NCBI
6.	Aw TC, Smith AB, Stephenson RL, et al: Occupational exposure to zeranol, an animal growth promoter. Br J Ind Med. 46:341–346. 1989.PubMed/NCBI
7.	Takemura H, Shim JY, Sayama K, Tsubura A, Zhu BT and Shimoi K: Characterization of the estrogenic activities of zearalenone and zeranol in vivo and in vitro. J Steroid Biochem Mol Biol. 103:17–7. 2007. View Article : Google Scholar : PubMed/NCBI
8.	Swan SH, Liu F, Overstreet JW, et al: Semen quality of fertile US males in relation to their mothers' beef consumption during pregnancy. Hum Reprod. 22:1497–1502. 2007.
9.	Linos E, Willett WC, Cho E, et al: Red meat consumption during adolescence among premenopausal women and risk of breast cancer. Cancer Epidemiol Biomarkers Prev. 17:2146–2151. 2008. View Article : Google Scholar : PubMed/NCBI
10.	Liu S and Lin YC: Transformation of MCF-10A human breast epithelial cells by zeranol and estradiol-17beta. Breast J. 10:514–521. 2004. View Article : Google Scholar : PubMed/NCBI
11.	Liu S, Sugimoto Y, Kulp SK, et al: Estrogenic down-regulation of protein tyrosine phosphatase gamma (PTP gamma) in human breast is associated with estrogen receptor alpha. Anticancer Res. 22:3917–3923. 2002.PubMed/NCBI
12.	Ye W, Xu P, Threlfall WR, et al: Zeranol enhances the proliferation of pre-adipocytes in beef heifers. Anticancer Res. 29:5045–5052. 2009.PubMed/NCBI
13.	Xu P, Ye W, Jen R, et al: Mitogenic activity of zeranol in human breast cancer cells is enhanced by leptin and suppressed by gossypol. Anticancer Res. 29:4621–4628. 2009.PubMed/NCBI
14.	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
15.	Ye W, Chang HL, Wang LS, et al: Modulation of multidrug resistance gene expression in human breast cancer cells by (-)-gossypol-enriched cottonseed oil. Anticancer Res. 27:107–116. 2007.PubMed/NCBI
16.	Darbre P, Yates J, Curtis S, et al: Effect of estradiol on human breast cancer cells in culture. Cancer Res. 43:349–354. 1983.PubMed/NCBI
17.	Yuri T, Tsukamoto R, Miki K, et al: Biphasic effects of zeranol on the growth of estrogen receptor-positive human breast carcinoma cells. Oncol Rep. 16:1307–1312. 2006.PubMed/NCBI
18.	Kuiper-Goodman T, Scott PM and Watanabe H: Risk assessment of the mycotoxin zearalenone. Regul Toxicol Pharmacol. 7:253–306. 1987. View Article : Google Scholar : PubMed/NCBI
19.	Giusti RM, Iwamoto K and Hatch EE: Diethylstilbestrol revisited: a review of the long-term health effects. Ann Intern Med. 122:778–788. 1995. View Article : Google Scholar : PubMed/NCBI
20.	Dinusson WE, Andrews FN and Beeson WM: The effects of stilbestrol, testosterone, thyroid alteration and spaying on the growth and fattening of beef heifers. J Anim Sci. 9:321–330. 1950.PubMed/NCBI
21.	Andrews FN, Beeson WM and Harper C: The effect of stilbestrol and testosterone on the growth and fattening of lambs. J Anim Sci. 8:578–582. 1949.
22.	Beral V and Colwell L: Randomised trial of high doses of stilboestrol and ethisterone in pregnancy: long-term follow-up of mothers. Br Med J. 281:1098–1101. 1980. View Article : Google Scholar : PubMed/NCBI
23.	Herbst AL, Ulfelder H and Poskanzer DC: Adenocarcinoma of the vagina. Association of maternal stilbestrol therapy with tumor appearance in young women. New Eng J Med. 284:878–881. 1971. View Article : Google Scholar
24.	Ellis PE, Maclean AB, Crow JC, et al: Expression of cyclin D1 and retinoblastoma protein in Paget's disease of the vulva and breast: an immunohistochemical study of 108 cases. Histopathology. 55:709–715. 2009.PubMed/NCBI
25.	Millar EK, Dean JL, McNeil CM, et al: Cyclin D1b protein expression in breast cancer is independent of cyclin D1a and associated with poor disease outcome. Oncogene. 28:1812–1820. 2009. View Article : Google Scholar : PubMed/NCBI
26.	Zwijsen RM, Buckle RS, Hijmans EM, et al: Ligand-independent recruitment of steroid receptor coactivators to estrogen receptor by cyclin D1. Genes Dev. 12:3488–3498. 1998. View Article : Google Scholar : PubMed/NCBI
27.	Toillon RA, Chopin V, Jouy N, et al: Normal breast epithelial cells induce p53-dependent apoptosis and p53-independent cell cycle arrest of breast cancer cells. Breast Cancer Res Treat. 71:269–280. 2002. View Article : Google Scholar : PubMed/NCBI
28.	Brugarolas J and Jacks T: Double indemnity: p53, BRCA and cancer. p53 mutation partially rescues developmental arrest in Brca1 and Brca2 null mice, suggesting a role for familial breast cancer genes in DNA damage repair. Nat Med. 3:721–722. 1997.
29.	Mandal S and Davie JR: Estrogen regulated expression of the p21 Waf1/Cip1 gene in estrogen receptor positive human breast cancer cells. J Cell Physiol. 224:28–32. 2010.PubMed/NCBI