Abstract
Immunohistochemistry using formalin-fixed, paraffin-embedded tissue, chromogen label, and light microscopy has traditionally been used to semiquantify estrogen receptor (ER) to guide diagnosis and management of breast cancer. Quantitation of ER for this purpose currently only assesses levels of the ER-alpha subtype. Considerable variability in results reported has been due to protocol and fixation variability, intraobserver and interobserver variability, and different scoring systems and thresholds for scoring ER positivity. Results can also vary with low expression levels of ER. ER-beta expression is reduced in breast and ovarian cancers and requires quantitation.
Herein we describe a novel approach to quantifying ERβ using older mouse ovarian surface epithelium, where ERβ is expressed at lower levels than ERα and is therefore harder to detect. We use an antibody highly specific to the ERβ1 isoform, together with immunofluorescence, confocal microscopy, and imaging and statistical software to achieve clear, reproducible, and unbiased quantitation of ERβ.
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References
Walter P, Green S, Greene G (1985) Cloning of the human estrogen receptor cDNA. Proc Natl Acad Sci U S A 82:7889–7893
White R, Lees JA, Needham M et al (1987) Structural organization and expression of the mouse estrogen receptor. Mol Endocrinol 1:735–744
Kuiper GG, Enmark E, Pelto-Huikko M et al (1996) Cloning of a novel receptor expressed in rat prostate and ovary. Proc Natl Acad Sci U S A 93:5925–5930
Mosselman S, Polman J, Dijkema R (1996) ER beta: identification and characterization of a novel human estrogen receptor. FEBS Lett 392:49–53
Tremblay GB, Tremblay A, Copeland NG et al (1997) Cloning, chromosomal localization, and functional analysis of the murine estrogen receptor beta. Mol Endocrinol 11:353–365
Gulliver LSM, Hurst PR (2011) Novel approaches to quantify loss of ERβ1 protein in older mouse ovarian surface epithelium: new tools to assess the role of ER protein subtypes in predisposing to ovarian epithelial cancer? Horm Cancer 2(4):204–213
Sas L, van Laere S, Dierick AM et al (2014) Study assessing the quality of quantification of estrogen receptor protein expression by immunohistochemistry and gene expression in breast cancer. Pathol Res Int 2014:372653. https://doi.org/10.1155/2014/372653
Zarella ER, Coulter M, Welsh AW et al (2016) Automated measurement of estrogen receptor in breast cancer: a comparison of fluorescent and chromogenic methods of measurement. Lab Invest 96:1016–1025
Hammond ME, Hayes DF, Dowsett M et al (2010) American Society of Clinical Oncology/College of American Pathologists guideline recommendations for immunohistochemical testing of estrogen and progesterone receptors in breast cancer. J Clin Oncol 28:2784–2795
Lazennec G (2006) Estrogen receptor beta, a possible tumor suppressor involved in ovarian carcinogenesis. Cancer Lett 231:151–157
Gulliver LSM, Hurst PR (2012) Repeat estradiol exposure differentially regulates protein expression patterns for estrogen receptor and E-cadherin in older mouse ovarian surface epithelium: implications for replacement and adjuvant hormonal therapies? Steroids 77(6):674–685
Srinivasan M, Sedmak D, Jewel S (2002) Effect of fixatives and tissue process on the content and integrity of nucleic acids. Am J Pathol 161:1961–1971
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Gulliver, L.S.M. (2019). In Vivo Quantitation of Estrogen Receptor β Subtype Expression in Ovarian Surface Epithelium Using Immunofluorescence Profiling and Confocal Microscopy. In: Badr, M. (eds) Nuclear Receptors. Methods in Molecular Biology, vol 1966. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9195-2_4
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DOI: https://doi.org/10.1007/978-1-4939-9195-2_4
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-9194-5
Online ISBN: 978-1-4939-9195-2
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