title: Lower HER-2/chromosome enumeration probe 17 ratio in cytologic HER-2 fluorescence in situ hybridization for breast cancers: three-dimensional analysis of intranuclear localization of centromere 17 and HER-2 signals.
abstract: BACKGROUND
  Fluorescence in situ hybridization (FISH) is the gold standard for assessing HER-2 status for breast cancers, and paraffin-embedded tissue sections are used routinely for HER-2 FISH. Cytologic samples also are used, but to the authors' knowledge, little is known regarding the reliability of these samples. The objective of this study was to elucidate the usefulness of cytologic specimens for HER-2 FISH testing.
  
  
  METHODS
  Histologic and cytologic specimens from 32 patients with invasive ductal carcinoma of the breast were subjected to comparative analysis of HER-2 status by FISH. FISH was performed by using a PathVysion HER-2 DNA Probe Kit according the manufacturer's instructions. The signal ratios of chromosome enumeration probe 17 (CEP17) and HER-2 were estimated and compared. In 15 cytologic specimens, the distance between signals (HER-2 and CEP17) and the nearest nuclear membrane were measured by using 3-dimensional image analysis and confocal microscopy.
  
  
  RESULTS
  Cytologic and histologic FISH results were compared. Signal ratios of HER-2/CEP17 were lower in cytologic specimens from 26 of 32 patients compared with the histologic material. Three-dimensional image analysis demonstrated that the distance between the CEP17 signal and the nuclear membrane was shorter than the distance between the HER-2 gene and the nuclear membrane.
  
  
  CONCLUSIONS
  The current results demonstrated that CEP17 could be lost more easily through histologic sectioning compared with the cytology results, because CEP17 is closer to the nuclear membrane. FISH analysis in cytologic specimens produced more accurate HER-2/CEP17 ratios, because the whole nucleus was subjected to FISH testing, compared with matched histologic specimens.
authors.affiliation: Division of Diagnostic Pathology, Department of Pathology, Tokai, University School of Medicine, Kanagawa, Japan. itoh-cyt@is.icc.u-tokai.ac.jp
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Itoh
authors.forename: Yoko
authors.initials: Y
authors.lastname: Miyajima
authors.forename: Shinobu
authors.initials: S
authors.lastname: Umemura
authors.forename: Robert Y
authors.initials: RY
authors.lastname: Osamura
mesh_terms: D001943:Breast Neoplasms / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics; D002467:Cell Nucleus / Q000235:genetics / Q000378:metabolism; D002503:Centromere / Q000235:genetics*; D002886:Chromosomes, Human, Pair 17 / Q000235:genetics*; D015342:DNA Probes / Q000235:genetics*; D005260:Female; D005784:Gene Amplification; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000235:genetics*
chemical_list: D002352:Carrier Proteins; D015342:DNA Probes; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human