Qualitative and quantitative differences in estrogen biotransformation in human breast glandular and adipose tissues: implications for studies using mammary biospecimens
Because of its assumed role in breast cancer etiology, estrogen biotransformation (and interaction of compounds therewith) has been investigated in human biospecimens for decades. However, little attention has been paid to the well-known fact that large inter-individual variations exist in the proportion of breast glandular (GLT) and adipose (ADT) tissues and less to adequate tissue characterization. To assess the relevance of this, the present study compares estrogen biotransformation in GLT and ADT. GLT and ADT were isolated from 47 reduction mammoplasty specimens derived from women without breast cancer and were characterized histologically and by their percentages of oil. Levels of 12 unconjugated and five conjugated estrogens were analyzed by GC- and UHPLC–MS/MS, respectively, and levels of 27 transcripts encoding proteins involved in estrogen biotransformation by Taqman® probe-based PCR. Unexpectedly, one-third of specimens provided neat GLT only after cryosection. Whereas 17β-estradiol, estrone, and estrone-3-sulfate were detected in both tissues, estrone-3-glucuronide and 2-methoxy-estrone were detected predominately in GLT and ADT, respectively. Estrogen levels as well as ratios 17β-estradiol/estrone and estrone-3-sulfate/estrone differed significantly between GLT and ADT, yet less than between individuals. Furthermore, estrogen levels in GLT and ADT correlated significantly with each other. In contrast, levels of most transcripts encoding enzymes involved in biotransformation differed more than between individuals and did not correlate between ADT and GLT. Thus, mixed breast tissues (and plasma) will not provide meaningful information on local estrogen biotransformation (and interaction of compounds therewith) whereas relative changes in 17β-estradiol levels may be investigated in the more abundant ADT.
KeywordsEstradiol Biotransformation Tissue levels Breast adipose tissue Breast glandular tissue
This work is part of the joint research project, IsoCross, entitled “Isoflavones: Cross-species comparison of metabolism, estrogen sensitivity, epigenetics and carcinogenesis”, which was supported in whole by the German Research Foundation to L. Lehmann (DFG LE 1329/10-1). The authors are indebted to Dr. Ulrike Waldhofen (mammoplasty specimen), Anne Scheffler (assistance during manuscript preparation), Ben Spielmann (help with sample collection, cryosection, microscopy and data administration), Sabine Winkler (help with analysis of estrogen conjugates), and Harald Schuchardt, Ersan Elemen, Thomas Kunz, Maryam Mahdiani (help with cryosection) as well as to Prof. Günter Vollmer (discussion of the manuscript).
Compliance with ethical standards
Conflict of interest
The authors have no conflict of interest to declare.
- Castagnetta LA, Granata OM, Traina A, Ravazzolo B, Amoroso M, Miele M, Bellavia V, Agostara B, Carruba G (2002) Tissue content of hydroxyestrogens in relation to survival of breast cancer patients. Clin Cancer Res 8:3146–3155Google Scholar
- Endogenous Hormones Breast Cancer Collaborative Group (2015) Steroid hormone measurements from different types of assays in relation to body mass index and breast cancer risk in postmenopausal women: Reanalysis of eighteen prospective studies. Steroids 99:49–55. https://doi.org/10.1016/j.steroids.2014.09.001 CrossRefGoogle Scholar
- Faupel-Badger JM, Fuhrman BJ, Xu X, Falk RT, Keefer LK, Veenstra TD, Hoover RN, Ziegler RG (2010) Comparison of liquid chromatography-tandem mass spectrometry, RIA, and ELISA methods for measurement of urinary estrogens. Cancer Epidemiol Biomarkers Prev 19:292–300. https://doi.org/10.1158/1055-9965.EPI-09-0643 CrossRefGoogle Scholar
- Ferlay J, Colombet M, Bray F (2018) Cancer Incidence in Five Continents, CI5plus: IARC CancerBase No. 9. Lyon, France: International Agency for Research on Cancer. http://ci5.iarc.fr. Last accessed 08 April 2019
- Figueroa JD, Pfeiffer RM, Patel DA, Linville L, Brinton LA, Gierach GL, Yang XR, Papathomas D, Visscher D, Mies C, Degnim AC, Anderson WF, Hewitt S, Khodr ZG, Clare SE, Storniolo AM, Sherman ME (2014) Terminal duct lobular unit involution of the normal breast: implications for breast cancer etiology. J Natl Cancer Inst 106:dju286. https://doi.org/10.1093/jnci/dju286 CrossRefGoogle Scholar
- Fleming JM, Miller TC, Quinones M, Xiao Z, Xu X, Meyer MJ, Ginsburg E, Veenstra TD, Vonderhaar BK (2010) The normal breast microenvironment of premenopausal women differentially influences the behavior of breast cancer cells in vitro and in vivo. BMC Med 8:27. https://doi.org/10.1186/1741-7015-8-27 CrossRefGoogle Scholar
- Gaikwad NW, Rogan EG, Cavalieri EL (2007) Evidence from ESI-MS for NQO1-catalyzed reduction of estrogen ortho-quinones. Free Radic Biol Med 43:1289–1298. https://doi.org/10.1016/j.freeradbiomed.2007.07.021 CrossRefGoogle Scholar
- Lee NA, Rusinek H, Weinreb J, Chandra R, Toth H, Singer C, Newstead G (1997) Fatty and fibroglandular tissue volumes in the breasts of women 20-83 years old: comparison of X-ray mammography and computer-assisted MR imaging. AJR Am J Roentgenol 168:501–506. https://doi.org/10.2214/ajr.168.2.9016235 CrossRefGoogle Scholar
- Lepine J, Bernard O, Plante M, Tetu B, Pelletier G, Labrie F, Belanger A, Guillemette C (2004) Specificity and regioselectivity of the conjugation of estradiol, estrone, and their catecholestrogen and methoxyestrogen metabolites by human uridine diphospho-glucuronosyltransferases expressed in endometrium. J Clin Endocrinol Metab 89:5222–5232. https://doi.org/10.1210/jc.2004-0331 CrossRefGoogle Scholar
- Nayeem F, Ju H, Brunder DG, Nagamani M, Anderson KE, Khamapirad T, Lu LJ (2014) Similarity of fibroglandular breast tissue content measured from magnetic resonance and mammographic images and by a mathematical algorithm. Int J Breast Cancer 2014:961679. https://doi.org/10.1155/2014/961679 CrossRefGoogle Scholar
- Savolainen-Peltonen H, Vihma V, Leidenius M, Wang F, Turpeinen U, Hamalainen E, Tikkanen MJ, Mikkola TS (2014) Breast adipose tissue estrogen metabolism in postmenopausal women with or without breast cancer. J Clin Endocrinol Metab 99:2661–2667. https://doi.org/10.1210/jc.2014-2550 CrossRefGoogle Scholar
- Sherman ME, Figueroa JD, Henry JE, Clare SE, Rufenbarger C, Storniolo AM (2012) The Susan G. Komen for the Cure Tissue Bank at the IU Simon Cancer Center: a unique resource for defining the “molecular histology” of the breast. Cancer Prev Res (Phila) 5:528–535. https://doi.org/10.1158/1940-6207.CAPR-11-0234 CrossRefGoogle Scholar
- Tsilidis KK, Allen NE, Key TJ, Dossus L, Lukanova A, Bakken K et al (2011) Oral contraceptive use and reproductive factors and risk of ovarian cancer in the European Prospective Investigation into cancer and nutrition. Br J Cancer 105:1436–1442. https://doi.org/10.1038/bjc.2011.371 CrossRefGoogle Scholar
- Vihma V, Wang F, Savolainen-Peltonen H, Turpeinen U, Hamalainen E, Leidenius M, Mikkola TS, Tikkanen MJ (2016) Quantitative determination of estrone by liquid chromatography-tandem mass spectrometry in subcutaneous adipose tissue from the breast in postmenopausal women. J Steroid Biochem Mol Biol 155(Pt A):120–125. https://doi.org/10.1016/j.jsbmb.2015.10.004 CrossRefGoogle Scholar
- Wang F, Vihma V, Badeau M, Savolainen-Peltonen H, Leidenius M, Mikkola T, Turpeinen U, Hamalainen E, Ikonen E, Wahala K, Fledelius C, Jauhiainen M, Tikkanen MJ (2012) Fatty acyl esterification and deesterification of 17 beta-estradiol in human breast subcutaneous adipose tissue. J Clin Endocrinol Metab 97:3349–3356. https://doi.org/10.1210/jc.2012-1762 CrossRefGoogle Scholar