title: Mutation analysis of the p53, APC, and p16 genes in the Barrett's oesophagus, dysplasia, and adenocarcinoma.
abstract: OBJECTIVE
  To study the loss of heterozygosity and the presence of mutations at the p53, p16/CDKN2, and APC genes in Barrett's oesophagus, low grade dysplastic oesophageal epithelium, and adenocarcinoma of the oesophagus; to relate the presence of alterations at these genes with the progression from Barrett's oesophagus to adenocarcinoma.
  
  
  METHODS
  DNA was extracted from paraffin blocks containing tissue from Barrett's oesophagus (12 samples), low grade dysplasia (15 cases), and adenocarcinoma (14 cases). Loss of heterozygosity (LOH) at the p53, p16, and APC genes was determined by comparing the autoradiographic patterns of several microsatellite markers between the normal tissue and the malignant tissue counterpart. SSCP was used to determine the presence of mutations at p53 (exons 5 to 8), p16 (exon 2), and APC. Homozygous deletion of the p16 gene was defined through polymerase chain reaction followed by Southern blot.
  
  
  RESULTS
  LOH at the p53, p16, and APC genes was not observed in Barrett's oesophagus without dysplasia, and increased to 90% (p53), 89% (p16), and 60% (APC) in the adenocarcinomas. The p53 gene was mutated in only two adenocarcinomas (codons 175 and 245). In one case a mutation at the APC gene (codon 1297) was found. No patient had mutation at the second exon of p16. However, this gene was homozygously deleted in three of the 12 adenocarcinomas.
  
  
  CONCLUSIONS
  The tumour suppressor genes p53, p16, and APC are often deleted in adenocarcinomas derived from Barrett's oesophagus. Mutations at these genes are also found in the adenocarcinomas, including the homozygous deletion of the p16 gene. However, the absence of genetic alterations in the Barrett's oesophagus and the low grade dysplastic epithelia suggest that mutations at these genes develop later in the progression from Barrett's oesophagus to adenocarcinoma.
authors.affiliation: Laboratorio de Genética Molecular-Servicio de Inmunología, Hospital Central de Asturias, Oviedo Spain.
authors.forename: M V
authors.initials: MV
authors.lastname: González
authors.forename: M L
authors.initials: ML
authors.lastname: Artímez
authors.forename: L
authors.initials: L
authors.lastname: Rodrigo
authors.forename: C
authors.initials: C
authors.lastname: López-Larrea
authors.forename: M J
authors.initials: MJ
authors.lastname: Menéndez
authors.forename: V
authors.initials: V
authors.lastname: Alvarez
authors.forename: R
authors.initials: R
authors.lastname: Pérez
authors.forename: M F
authors.initials: MF
authors.lastname: Fresno
authors.forename: M J
authors.initials: MJ
authors.lastname: Pérez
authors.forename: A
authors.initials: A
authors.lastname: Sampedro
authors.forename: E
authors.initials: E
authors.lastname: Coto
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics*; D001471:Barrett Esophagus / Q000235:genetics* / Q000473:pathology; D004252:DNA Mutational Analysis; D018450:Disease Progression; D004938:Esophageal Neoplasms / Q000235:genetics*; D016368:Frameshift Mutation; D017353:Gene Deletion; D017491:Genes, APC / Q000235:genetics; D016147:Genes, Tumor Suppressor / Q000235:genetics*; D016158:Genes, p53 / Q000235:genetics; D006579:Heterozygote; D006801:Humans; D017354:Point Mutation; D017422:Sequence Analysis, DNA