title: p16INK4a promoter is hypermethylated at a high frequency in esophageal adenocarcinomas.
abstract: Loss of heterozygosity (LOH) of 9p21, which contains the p16INK4a tumor suppressor gene locus, is one of the most frequent genetic abnormalities in human neoplasia, including esophageal adenocarcinomas. Only a minority of Barrett's adenocarcinomas with 9p21 LOH have a somatic mutation in the remaining p16 allele, and none have been found to have homozygous deletions. To determine whether p16 promoter hypermethylation may be an alternative mechanism for p16 inactivation in esophageal adenocarcinomas, we examined the methylation status of the p16 promoter in flow-sorted aneuploid cell populations from 21 patients with premalignant Barrett's epithelium or esophageal adenocarcinoma. Using bisulfite modification, primer-extension preamplification, and methylation-specific PCR, we demonstrate that the methylation assay can be performed on 2 ng of DNA (approximately 275 cells). Eight of 21 patients (38%) had p16 promoter hypermethylation and 9p21 LOH, including 3 patients who had only premalignant Barrett's epithelium. Our data suggest that promoter hypermethylation with LOH is a common mechanism for inactivation of p16 in the pathogenesis of esophageal adenocarcinomas.
authors.affiliation: Division of Public Health Sciences, Program in Molecular and Cellular Biology, Fred Hutchinson Cancer Research Center, Seattle, Washington 98104, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Wong
authors.forename: M T
authors.initials: MT
authors.lastname: Barrett
authors.forename: R
authors.initials: R
authors.lastname: Stöger
authors.forename: M J
authors.initials: MJ
authors.lastname: Emond
authors.forename: B J
authors.initials: BJ
authors.lastname: Reid
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics*; D001471:Barrett Esophagus / Q000235:genetics*; D002352:Carrier Proteins / Q000235:genetics*; D002872:Chromosome Deletion; D002899:Chromosomes, Human, Pair 9 / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16; D019175:DNA Methylation; D004938:Esophageal Neoplasms / Q000235:genetics*; D016147:Genes, Tumor Suppressor; D006579:Heterozygote; D006801:Humans; D011401:Promoter Regions, Genetic; D014407:Tumor Cells, Cultured
chemical_list: D002352:Carrier Proteins; D019941:Cyclin-Dependent Kinase Inhibitor p16