Detection of genetic changes in Barrett's adenocarcirioma and Barrett's esophagus by DNA in situ hybridization and immunohistochemistry

Abstract

We have investigated the occurrence of chromosomal DNA and cell cycle‐related protein changes in Barrett's epithelium and adenocarcinoma. The presence of numerical chromosomal aberrations was studied by applying nonisotopic in situ hybridization (ISH) with (peri‐)centromeric DNA probes, specific for chromosomes 7, 8, 17, and Y, to routinely processed tissue sections of five cases (4 male, 1 female) of Barrett's adenocarcinoma and adjacent Barrett's epithelium. Cell cycle‐related protein expression was studied by immunohistochemistry (IHC) for p53 protein and the Ki‐67 antigen (Mib‐1) in subsequent sections. P53 protein overexpression was found in 3 of the 5 tumors. Overrepresentation of chromosome 8 and loss of chromosome 17 were found in 2 adenocarcinomas, both also negative for p53 protein overexpression. Y‐loss, mostly clonal, was detected in 3 of the 4 male adenocarcinomas and 2 cases of adjacent Barrett's epithelium. One tumor had both areas of overrepresentation and loss of the Y chromosome. All Barrett's adenocarcinomas appeared to contain aneuploid cell populations. No relation was found between cell proliferation characteristics and chromosomal aberrations. We conclude that ISH with chromosome specific DNA probes can be applied for the assessment of potentially important numerical chromosome changes in Barrett's esophagus. Further, the combination of IHC and ISH is useful for evaluation of specific genetic events. © 1994 Wiley‐Liss, Inc.

Chromosomal gains and genomic loss of p53 and p16 genes in Barrett's esophagus detected by fluorescence in situ hybridization of cytology specimens

“…[34][35][36][37][38][39] The studies consistently show an accumulation of chromosomal abnormalities as the histologic changes progress from intestinal metaplasia to dysplasia and carcinoma. 4,[40][41][42][43] Previously reported changes detectable by FISH in esophagectomy specimens suggested that gains of chromosomes 6, 7, 11, and 12 might be among the numerical abnormalities occurring most frequently in the Barrett's esophagus-associated HGD and adenocarcinoma, findings which we have confirmed with the use of endoscopic cytology specimens. 22 Only sparse data are available in the literature regarding the use of FISH on endoscopic brush cytology specimens for biomarker assessment in Barrett's esophagus.…”

“…The loss of cell cycle check point control, which results from this alteration, can lead to asymmetrical chromosome segregation during cell division and subsequent aneuploidy. 40 Mutations of p53 and 17p13.1 loss of heterozygosity have been reported in upto 92 and 100% of esophageal adenocarcinomas. [41][42][43] Our study demonstrates that the genomic loss of the 17p13.1 region (p53 gene) occurs in the majority of the HGD/ adenocarcinoma cases (76%).…”

“…To determine if there are specific genetic changes in Barrett's esophagus with associated high-grade dysplasia/intramucosal adenocarcinoma compared to those without dysplasia, we performed fluorescence in situ hybridization (FISH) on cytologic specimens using probes to chromosomes and genomic regions previously described as altered in this disease. We studied archival brush cytology slides from 40 Barrett's esophagus patients: 21 with biopsy-proven high-grade dysplasia/carcinoma and 19 with no dysplasia and a minimum 5 years of negative follow-up. Centromeric enumeration probes (CEP) for chromosomes 6, 7, 11, and 12, and locus-specific probes (LSI) for 9p21 (p16 gene), and 17p13.1 (p53 gene) loci along with their corresponding CEP (9 and 17, respectively) were used in this study.…”

Interphase cytogenetics of prostatic adenocarcinoma and precursor lesions: Analysis of 25 radical prostatectomies and 17 adjacent prostatic intraepithelial neoplasias

“…Internal controls (normal prostate glands: 13 cases; BPH: 4 cases; leukocytes: 131 cases; other cells: 5 cases) on the same tissue sections always showed a diploid pattern (van Dekken and Alers, 1993;Krishnadath et al, 1994). T h e number of nuclei with a hyperdiploid spot number (likely artifacts) in these internal controls never exceeded 2.5%.…”

Chromosomal analysis of Barrett's cells: demonstration of instability and detection of the metaplastic lineage involved

“…15,34,35 But, it remains to clarify its role as a cause or an effect. So far, no specific tumor suppressor genes have been mapped to the Y chromosome and Krishnadath et al, 36 who found no correlation between proliferation rates as detected by Ki-67 and loss of Y chromosome, did not confirm the hypothesis that its loss might confer proliferative advantage. Despite its unclear role as a side effect or a true initiating mechanism, all previous studies described chromo- some Y loss in Barrett's esophagus adjacent to neoplasia.…”