Special in situ techniques sectionProgress in pathologyFluorescence in situ hybridization in diagnostic cytology☆
Introduction
For more than a half century, the Papanicolaou stain and other cytochemical stains have been the primary modality used to identify tumor cells in cytologic specimens. However, in recent years, ancillary techniques such as DNA ploidy analysis by flow cytometry/image cytometry, immunocytology, and fluorescence in situ hybridization (FISH) have been increasingly used to facilitate the identification of neoplastic cells in cytologic specimens.
It is now widely recognized that tumors result from genetic and epigenetic alterations that activate oncogenes and inactivate tumor suppressor genes. Mutations within tumors occur at both the chromosomal and molecular level. Types of chromosomal alterations that occur in tumors include aneuploidy, deletion, amplification, and translocation.
FISH is a technique that uses fluorescently labeled DNA probes to detect chromosomal alterations in cells. FISH can detect various types of cytogenetic alterations including aneusomy (ie, abnormalities of chromosome copy number), duplication, amplification, deletion, and translocation. There are 2 types of FISH probes: chromosome enumeration probes (CEPs) and locus-specific indicator (LSI) probes. CEPs are used to detect aneusomy, whereas LSI probes are generally used to detect deletion, duplication, or amplification of specific genes.
Because tumor cells generally contain chromosomal alterations, FISH should be able to detect cells that have chromosomal abnormalities consistent with neoplasia in exfoliative and aspiration cytology specimens. This has proven to be the case, and FISH is now used to detect bladder and biliary tract cancers in urine and biliary tract brushings [1], [2], respectively, and will likely soon be used to identify tumor cells in a variety of other cytologic specimens. The remainder of this review will discuss how FISH has been used to identify tumor cells in various types of cytologic specimens.
Section snippets
FISH for bladder cancer detection
In the mid- to late 1990s, data began to appear in the literature suggesting that FISH might be a useful test for detecting bladder cancer in voided urine and bladder washing specimens [3], [4], [5], [6], [7]. In 2000, Sokolova et al [8] introduced what would become the first commercially available FISH probe set for bladder cancer detection in voided urine. This 4-target, multicolor FISH probe set, UroVysion (Abbott Molecular Inc, Des Plaines, IL), contains CEP probes for chromosomes 3, 7, and
FISH for the detection of biliary tract malignancy
Strictures of the extrahepatic bile ducts are a common occurrence in clinical practice. Although these strictures are often due to malignancies such as cholangiocarcinoma or pancreatic cancer, they can be the result of nonmalignant etiologies such as choledocholithiasis, chronic pancreatitis, surgical trauma, and ischemia. The distinction between benign and malignant pancreatobiliary strictures can be difficult because a cancer may be present but not identified on cross-sectional imaging
FISH for the detection of lung cancer
In 2007, approximately 213 000 people in the United States will be diagnosed with lung cancer, and 160 000 will die from their disease [39]. The dismal 5-year survival of 15% for all patients diagnosed with lung cancer has been attributed to the fact that most patients have advanced disease at the time of diagnosis [39], [40], [41][41]. Cytology (brushings and washings) and biopsy specimens collected during bronchoscopy are widely used techniques to diagnose malignancy in patients suspected of
FISH for the detection of dysplasia and adenocarcinoma in patients with Barrett's esophagus
Barrett's esophagus is a preneoplastic condition in which the squamous epithelium of the distal esophagus undergoes transformation to intestinal metaplasia. Patients with Barrett's esophagus are approximately 125 times more likely to develop carcinoma than the general population and therefore must undergo regular surveillance [53]. Recent studies suggests that FISH has the potential to be used to identify dysplasia (low- and high-grade) and adenocarcinoma in patients with Barrett's esophagus
Summary
Studies performed over the past 5 to 10 years indicate that FISH represents a promising new way of detecting tumor cells in cytologic specimens. Instead of using a cytochemical stain to look for morphological features characteristic of neoplasia, FISH examines for chromosomal alterations that are consistent with a diagnosis of neoplasia. Most studies have shown that FISH has significantly higher sensitivity than conventional cytology for the detection of tumor cells in most specimen types.
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Cited by (0)
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Dr Halling receives grant funding from Abbott Molecular, Inc (Des Plaines, IL) to develop fluorescence in situ hybridization assays for the detection of neoplastic cells in cytologic specimens. He holds a patent on the UroVysion probe set (Abbott Molecular, Inc) and has filed patents for the lung cancer and Barrett's probe sets.