Microdissection and molecular genetic analysis of HER2/neu in breast carcinoma Journal Article


Authors: Youngson, B. J.; Anelli, A.; Van Zee, K. J.; Borgen, P. I.; Norton, L.; Rosen, P. P.
Article Title: Microdissection and molecular genetic analysis of HER2/neu in breast carcinoma
Abstract: Precise correlation of histomorphology with molecular genetic analysis is difficult in tissues composed of heterogeneous cell populations. We describe here a novel microdissection technique employed to correlate HER2/neu (HER2) immunohistochemical staining with HER2 genetic analysis in formalin-fixed, paraffin-embedded breast tissue. Fourteen invasive ductal carcinomas were selected from the pathology files of Memorial Sloan-Kettering Cancer Center that had been immunostained for HER2. Seven tumors showed typical membrane immunoreactivity and seven were negative. A dissecting microscope was then used to isolate minute (≤1 mm x 1 mm) areas of invasive carcinoma and normal breast tissue for molecular study. To document the type of cell sample submitted for polymerase chain reaction (PCR) analysis, each microdissected piece of tissue was photographed prior to removal from the glass slide. A preliminary study of four cases compared the results of PCR and genetic analysis using microdissected hematoxylin and eosin (H and E)-stained tissue, unstained dewaxed tissue, and destained dewaxed tissue in four specimens. Similar results were obtained with all three tissue preparations. Thereafter, H and E stained sections were selected as the tissue preparation of choice because tissue details were seen more clearly. There was complete correlation of immunohistochemical staining and HER2 analysis by PCR in all 14 cases. In the final 10 cases, the PCR product was resolved by gel electrophoresis and quantified by optical densitometry. Fourfold to eightfold amplification of HER2 was found in the five tumor specimens that immunohistochemically stained for HER2. A single copy of HER2 was found in all HER2-negative tumors and in normal breast tissue. We conclude that it is possible to quantify gene amplification of HER2 in minute samples of H and E-stained normal and malignant breast tissue. This micro-dissection technique can be applied to correlative histologic-molecular genetic analysis in a wide variety of tumor types.
Keywords: immunohistochemistry; controlled study; unclassified drug; human cell; polymerase chain reaction; proto oncogene; epidermal growth factor receptor 2; oncogenes; breast carcinoma; densitometry; invasive carcinoma; eosin; hematoxylin; intraductal carcinoma; microdissection; gel electrophoresis; her2/neu; human; article
Journal Title: American Journal of Surgical Pathology
Volume: 19
Issue: 12
ISSN: 0147-5185
Publisher: Lippincott Williams & Wilkins  
Date Published: 1995-12-01
Start Page: 1354
End Page: 1358
Language: English
DOI: 10.1097/00000478-199512000-00002
PUBMED: 7503357
PROVIDER: scopus
DOI/URL:
Notes: Article -- Export Date: 28 August 2018 -- Source: Scopus
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MSK Authors
  1. Larry Norton
    758 Norton
  2. Kimberly J Van Zee
    293 Van Zee
  3. Patrick I Borgen
    253 Borgen
  4. Paul P Rosen
    201 Rosen