Measurement of markers for breast cancer in a model system using laser scanning cytometry

• Published in: Cytometry (New York, N.Y.) N.Y.), 2000-11, Vol.41 (3), p.166-171
• Main Authors: Zabaglo, Lila, Ormerod, Michael G., Dowsett, Mitch
• Format: Article
• Language: English
• Subjects: Apoptosis; breast cancer; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Carcinoma - metabolism; Carcinoma - pathology; Estradiol - analogs & derivatives; Estradiol - pharmacology; Estrogen Antagonists - pharmacology; Female; fine needle aspirates; Humans; Immunohistochemistry; Ki-67 Antigen - immunology; Ki-67 Antigen - metabolism; Ki67; laser scanning cytometry; Microscopy, Confocal - methods; Microscopy, Fluorescence; PgR; Receptors, Estrogen - drug effects; Receptors, Estrogen - immunology; Receptors, Estrogen - metabolism; Receptors, Progesterone - immunology; Receptors, Progesterone - metabolism; Tumor Cells, Cultured
• ISSN: 0196-4763; 1097-0320
• DOI: 10.1002/1097-0320(20001101)41:3<166::AID-CYTO2>3.0.CO;2-Y

Background A variety of markers, including Ki67, estrogen receptors (ER), and progesterone receptors (PgR), are frequently measured in fine needle aspirates (FNA) from human breast carcinomas. We used a human breast carcinoma cell line, MCF7, as a model system to investigate the use of laser scanning cytometry (LSC) for the measurement of these markers. Additionally, we measured the number of apoptotic cells. Methods Cells were treated with drugs to vary the expression of markers and the number of apoptotic cells. They were then fixed on microscope slides. For LSC, the cells were stained for the different markers with fluorescein using immunofluorescence and for apoptotic cells using the TUNEL assay. The nuclei were counterstained with propidium iodide. A parallel set of slides was stained using horseradish peroxidase and diaminobenzidine and scored manually by conventional light microscopy. Results The results from the LSC closely paralleled those obtained by manual scoring of immunohistochemical stains. Conclusions It should be possible to use LSC for the routine measurement of nuclear markers in FNAs from human breast carcinomas. Cytometry 41:166–171, 2000 © 2000 Wiley‐Liss, Inc.