Sequential Colocalization of ERa, PR, and AR Hormone Receptors Using Confocal Microscopy Enables New Insights into Normal Breast and Prostate Tissue and Cancers

• Published in: Cancers 2020-11, Vol.12 (12), p.3591
• Main Authors: Chenlo, Miguel, Aliyev, Elvin, Rodrigues, Joana S, Vieiro-Balo, Paula, Blanco Freire, Manuel N, Cameselle-Teijeiro, José Manuel, Alvarez, Clara V
• Format: Article
• Language: English
• Subjects: Antibodies; Automation; Breast cancer; Confocal microscopy; Epithelium; Immunohistochemistry; Light microscopy; Localization; Microscopy; Nuclear receptors; Pathology; Prostate; Prostate cancer; Stains & staining
• ISSN: 2072-6694; 2072-6694
• DOI: 10.3390/cancers12123591

Multiplex immunohistochemistry (mIHC) use markers staining different cell populations applying widefield optical microscopy. Resolution is low not resolving subcellular co-localization. We sought to colocalize markers at subcellular level with antibodies validated for clinical diagnosis, including the single secondary antibody (combination of anti-rabbit/mouse-antibodies) used for diagnostic IHC with any primary antibody, and confocal microscopy. We explore colocalization in the nucleus ( ) of nuclear hormone receptors (ERa, PR, and AR) along with the baseline marker p63 in paired samples of breast and prostate tissues. We established ColNu mIHCF as a reliable technique easily implemented in a hospital setting. In ERa+ breast cancer, we identified different colocalization patterns (nuclear or cytoplasmatic) with PR and AR on the luminal epithelium. A triple-negative breast-cancer case expressed membrane-only ERa. A PR-only case was double positive PR/p63. In normal prostate, we identified an ERa+/p63+/AR-negative distinct population. All prostate cancer cases characteristically expressed ERa on the apical membrane of the AR+ epithelium. We confirmed this using ERa IHC and needle-core biopsies. is feasible and already revealed a new marker for prostate cancer and identified sub-patterns in breast cancer. It could be useful for pathology as well as for functional studies in normal prostate and breast tissues.