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Prepubertal physical activity up-regulates estrogen receptor β, BRCA1 and p53 mRNA expression in the rat mammary gland

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Abstract

Findings in BRCA1 mutation carriers suggest that physical activity, particularly during childhood, may be linked to a reduced risk of developing breast cancer. We investigated whether physical activity at puberty alters the expression of BRCA1 and two other tumor suppressor genes—p53 and estrogen receptor (ER)-β—in rats. In addition, the effects on ER-α expression, mammary proliferation and functional epithelial differentiation were investigated as markers of altered mammary cancer risk in rats exposed to regular physical activity at puberty. Female Sprague Dawley rat pups were randomized to voluntary exercise, sham-exercise control and non-manipulated control groups. Treadmill training (20–25 m/min, 15% grade, 30 min/day, 5 days/week) started on postnatal day 14 and continued through day 32. Third thoracic mammary glands (n = 5 per group and age) were obtained at days 32, 48 and 100 and assessed for changes in morphology through wholemounts, and at 100 days cell proliferation by using Ki67 staining, protein levels of ER-α and ER-β by immunohistochemistry, and mRNA expression levels of BRCA1, p53, ER-α and ER-β by real-time PCR. Mammary glands of rats exposed to exercise during puberty contained fewer terminal end buds (TEBs) and a higher number of differentiated alveolar buds and lobules than the sham controls. However, cell proliferation was not significantly altered among the groups. ER-α protein levels were significantly reduced, while ER-β levels were increased in the mammary ducts and lobular epithelial structures of 100-day old rays which were voluntarily exercised at puberty, compared to sham controls. ER-β, BRCA1 and p53 mRNA levels were significantly higher in the mammary glands of 100-day-old exercised versus sham control rats. Pubertal physical activity reduced mammary epithelial targets for neoplastic transformation through epithelial differentiation and it also up-regulated tumor suppressor genes BRCA1, p53 and ER-β, and reduced ER-α/ER-β ratio in the mammary gland. It remains to be determined whether the up-regulation of BRCA1, and perhaps p53, explains the protective effect of childhood physical activity against breast cancer in women who carry a germline mutation in one of the BRCA1 alleles.

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Acknowledgments

We thank Dr. Elizabeth Cho-Fertick who provided medical writing services, funded by National Cancer Institute (U54 CA00100971 for L.H.-C.). The study was funded by grants from Prevent Cancer Foundation (K·W.) and National Cancer Institute (U54 CA00100971, L.H.-C.).

Authors’ contributions

The work described in the manuscript was designed to test a hypothesis proposed by Dr. Leena Hilakivi-Clarke who provided overall direction for the study. Drs. Kim Westerlind and Robert Strange performed the animal study and provided tissues for the analysis, Dr. Mingyue Wang did most of the gene and protein expression experiments, together with Drs. Bin Yu, Galam Khan and Dipti Patil. Graduate student Kelly Boeneman processed the mammary glands and performed morphological assessment.

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Authors and Affiliations

  1. Department of Oncology, Georgetown University, Washington, DC, USA

    M. Wang, B. Yu, G. Khan, D. Patil, K. Boeneman & L. Hilakivi-Clarke

  2. Division of Endocrinology, University of Colorado Health Sciences Center, Denver, CO, USA

    K. Westerlind

  3. Division of Medical Oncology, University of Colorado Health Sciences Center, Denver, CO, USA

    R. Strange

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  1. M. Wang
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Correspondence to L. Hilakivi-Clarke.

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Wang, M., Yu, B., Westerlind, K. et al. Prepubertal physical activity up-regulates estrogen receptor β, BRCA1 and p53 mRNA expression in the rat mammary gland. Breast Cancer Res Treat 115, 213–220 (2009). https://doi.org/10.1007/s10549-008-0062-x

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