The following antibodies were used for immunostaining: 1D5 (Dako, Glostrup, Denmark) for ER, PgR636 (Dako) for progesterone receptor (PgR) and Hercep Test (Dako) for HER2. For evaluation of HER2 gene amplification, dual in situ hybridisation (DISH) was performed with INFORM HER2 Dual ISH DNA Probe Cocktail assay (Ventana Medical Systems, Inc., Tuscon, AZ, USA). ER, PgR and HER2 expression were evaluated in accordance with the American Society of Clinical Oncology and College of American Pathologists (11 (link),12 (link)) criteria. In addition, the degree of ER and PgR staining ≥1% and the specimen was determined as positive. The proportional scores of cells membrane HER2 staining intensity were as follows: scores 0, 1+, 2+ and 3+. HER2 immunostaining with a score of 2+ was subjected to a DISH assay to assess the gene amplification of HER2. A HER2 score of 2+/DISH positive or 3+ was defined as HER2-positive cancer. Patients with ER-positive and/or PgR-positive breast cancer were defined as a hormonal receptor (HR)-positive breast cancer.
Protocol detail
Immunostaining and HER2 Evaluation in Breast Cancer
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Antibodies
Assay
Breast cancer
Cancer
Dna probe
Her2
In situ hybridisation
Pathologists
Patients
Progesterone receptor
Corresponding Organization :
Other organizations : Saitama Cancer Center, Gunma University, International University of Health and Welfare
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Variable analysis
independent variables
- Antibodies used for immunostaining: 1D5 (Dako, Glostrup, Denmark) for ER, PgR636 (Dako) for progesterone receptor (PgR), and Hercep Test (Dako) for HER2
- Dual in situ hybridisation (DISH) assay with INFORM HER2 Dual ISH DNA Probe Cocktail (Ventana Medical Systems, Inc., Tuscon, AZ, USA) for evaluation of HER2 gene amplification
- ER expression
- PgR expression
- HER2 expression
- HER2 gene amplification
- ER, PgR, and HER2 expression evaluated according to the American Society of Clinical Oncology and College of American Pathologists criteria
- Specimen determined as ER and/or PgR positive if the degree of staining is ≥1%
- HER2 immunostaining with a score of 2+ subjected to DISH assay to assess gene amplification
- HER2 score of 2+/DISH positive or 3+ defined as HER2-positive cancer
- Patients with ER-positive and/or PgR-positive breast cancer defined as hormonal receptor (HR)-positive breast cancer
- None specified
- None specified
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