ERβ splice variant expression in four large cohorts of human breast cancer patient tumors

Hallie Wimberly, Gang Han, Dushanthi Pinnaduwage, Leigh C. Murphy, Xiaohong Rose Yang, Irene L. Andrulis, Mark E. Sherman, Jonine Figueroa, David L. Rimm

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Though the role of Estrogen Receptor (ER)α in breast cancer has been studied extensively, there is little consensus about the role of alternative ER isoform ERβ in breast cancer biology. ERβ has significant sequence homology to ERα but is located on a different chromosome and maintains both overlapping and unique functional attributes. Five variants exist, resulting from alternative splicing of the C-terminal region of ERβ. The relevance of ERβ variants in breast cancer outcomes and response to therapy is difficult to assess because of conflicting reports in the literature, likely due to variable methods used to assess ERβ in patient tumors. Here, we quantitatively assess expression of ERβ splice variants on over 2,000 breast cancer patient samples. Antibodies against ERβ variants were validated for staining specificity in cell lines by siRNA knockdown of ESR2 and staining reproducibility on formalin-fixed paraffin-embedded tissue by quantitative immunofluorescence (QIF) using AQUA technology. We found antibodies against splice variants ERβ1 and ERβ5, but not ERβ2/cx, which were sensitive, specific, and reproducible. QIF staining of validated antibodies showed both ERβ1 and ERβ5 QIF scores, which have a normal (bell shaped) distribution on most cohorts assessed, and their expression is significantly associated with each other. Extensive survival analyses show that ERβ1 is not a prognostic or predictive biomarker for breast cancer. ERβ5 appears to be a context-dependent marker of worse outcome in HER2-positive and triple-negative patients, suggesting an unknown biological function in the absence of ERα.

Original languageEnglish (US)
Pages (from-to)657-667
Number of pages11
JournalBreast Cancer Research and Treatment
Volume146
Issue number3
DOIs
StatePublished - 2014

Fingerprint

Estrogen Receptors
Breast Neoplasms
Neoplasms
Fluorescent Antibody Technique
Staining and Labeling
Antibodies
Alternative Splicing
Survival Analysis
Sequence Homology
Paraffin
Formaldehyde
Small Interfering RNA
Protein Isoforms
Chromosomes
Biomarkers
Technology
Cell Line

Keywords

  • Antibody validation
  • Breast cancer prognosis and prediction
  • Estrogen receptors
  • Quantitative immunofluorescence
  • Triple-negative breast cancer

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Wimberly, H., Han, G., Pinnaduwage, D., Murphy, L. C., Yang, X. R., Andrulis, I. L., ... Rimm, D. L. (2014). ERβ splice variant expression in four large cohorts of human breast cancer patient tumors. Breast Cancer Research and Treatment, 146(3), 657-667. https://doi.org/10.1007/s10549-014-3050-3

ERβ splice variant expression in four large cohorts of human breast cancer patient tumors. / Wimberly, Hallie; Han, Gang; Pinnaduwage, Dushanthi; Murphy, Leigh C.; Yang, Xiaohong Rose; Andrulis, Irene L.; Sherman, Mark E.; Figueroa, Jonine; Rimm, David L.

In: Breast Cancer Research and Treatment, Vol. 146, No. 3, 2014, p. 657-667.

Research output: Contribution to journalArticle

Wimberly, H, Han, G, Pinnaduwage, D, Murphy, LC, Yang, XR, Andrulis, IL, Sherman, ME, Figueroa, J & Rimm, DL 2014, 'ERβ splice variant expression in four large cohorts of human breast cancer patient tumors', Breast Cancer Research and Treatment, vol. 146, no. 3, pp. 657-667. https://doi.org/10.1007/s10549-014-3050-3
Wimberly, Hallie ; Han, Gang ; Pinnaduwage, Dushanthi ; Murphy, Leigh C. ; Yang, Xiaohong Rose ; Andrulis, Irene L. ; Sherman, Mark E. ; Figueroa, Jonine ; Rimm, David L. / ERβ splice variant expression in four large cohorts of human breast cancer patient tumors. In: Breast Cancer Research and Treatment. 2014 ; Vol. 146, No. 3. pp. 657-667.
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