Chromogenic in situ hybridization - A practical alternative for fluorescence in situ hybridization to detect HER-2/neu oncogene amplification in archival breast cancer samples
M. Tanner et al., Chromogenic in situ hybridization - A practical alternative for fluorescence in situ hybridization to detect HER-2/neu oncogene amplification in archival breast cancer samples, AM J PATH, 157(5), 2000, pp. 1467-1472
Citations number
16
Categorie Soggetti
Research/Laboratory Medicine & Medical Tecnology","Medical Research Diagnosis & Treatment
Determination of HER-2/neu oncogene amplification has become necessary for
selection of breast cancer patients for trastuzumab (Herceptin) therapy. Fl
uorescence in situ hybridization (FISH) is currently regarded as a gold sta
ndard method for detecting HER2/neu amplification, but it is not very pract
ical for routine histopathological laboratories. We evaluated a new modific
ation of in situ hybridization, the chromogenic ill situ hybridization (CIS
H), which enables detection of HER-2/neu gene copies with conventional pero
xidase reaction. Archival formalin-fixed paraffin-embedded tumor tissue sec
tions were pretreated (by heating In a microwave oven and using enzyme dige
stion) and hybridized with a digoxigenin-labeled DNA probe. The probe was d
etected with antidigoxigenin fluorescein, anti-fluorescein peroxidase, and
diaminobenzidine. Gene copies visualized by CISH could be easily distinguis
hed with a x40 objective in hematoxylin-stained tissue sections. HER-2/neu
amplification typically appeared as large peroxidase-positive intranuclear
gene copy clusters. CISH and FISH (according to Vysis, made from frozen pul
verized tumor samples) correlated well in a series of 157 breast cancers (k
appa coefficient, 0.81). The few different classifications were mostly beca
use of low-level amplifications by FISH that were negative by CISH and immu
nohistochemistry with monoclonal antibody CB-11, We conclude that CISH, usi
ng conventional bright-field microscopy in evaluation, is a useful alternat
ive for determination of HER-2/neu amplification in paraffin-embedded tumor
samples, especially for confirming the immunohistochemical staining result
s.