title: Molecular detection of early-stage laryngopharyngeal squamous cell carcinomas.
abstract: OBJECTIVE
  We sought to determine whether early-stage laryngopharyngeal squamous cell carcinomas (SCC) can be detected through molecular analysis of exfoliated cells collected with the use of a pharyngoesophageal brush (PEB).
  
  
  METHODS
  Thirty-three patients with a single, untreated, early-stage (T1 or T2) SCC of the supraglottic larynx or pharynx underwent collection of cells with a PEB, followed by endoscopic biopsy of the tumor. PEB specimens were also collected from five healthy subjects. PEB samples and tumor tissue were examined for hypermethylation of p16INK4a (CDKN2) gene promoter CpG islands (assayed by methylation-specific PCR) and UT5085 tetranucleotide microsatellite instability (assayed by GeneScan analysis). PEB samples were also subjected to cytologic analysis.
  
  
  RESULTS
  Eight of 33 (24%) tumors exhibited a bandshift at UT5085, and 14 of 33 (42%) exhibited hypermethylation at the p16 promoter. Overall, 17 of 33 (52%) patients had at least one of the two markers in their tumor. Cytologic analysis of PEB samples revealed tumor in 4 of 33 (12%) patients; cytologic findings were normal in all five control subjects. Molecular analysis of PEB samples revealed tumor DNA in 13 of 17 (76%) patients with at least one of the two molecular markers in their tumor. Eight of 14 (57%) patients with p16 hypermethylation in their tumor and 8 of 8 (100%) patients with UT5085 microsatellite instability in their tumor had similar findings in the PEB samples. None of the PEB samples from the control subjects or patients with neither molecular marker in their tumor displayed abnormality.
  
  
  CONCLUSIONS
  Molecular analysis of PEB samples holds promise for the early detection of early-stage laryngopharyngeal SCCs. New molecular markers need to be identified to increase the sensitivity of molecular screening.
authors.affiliation: Department of Head and Neck Surgery, Institut Gustave-Roussy, Villejuif, France.
authors.forename: Stephane
authors.initials: S
authors.lastname: Temam
authors.forename: Jean
authors.initials: J
authors.lastname: Bénard
authors.forename: Christelle
authors.initials: C
authors.lastname: Dugas
authors.forename: Martine
authors.initials: M
authors.lastname: Trassard
authors.forename: Emmanuelle
authors.initials: E
authors.lastname: Gormally
authors.forename: Jean-Charles
authors.initials: JC
authors.lastname: Soria
authors.forename: Sandrine
authors.initials: S
authors.lastname: Faivre
authors.forename: Bernard
authors.initials: B
authors.lastname: Luboinski
authors.forename: Patrick
authors.initials: P
authors.lastname: Marandas
authors.forename: Pierre
authors.initials: P
authors.lastname: Hainaut
authors.forename: Gilbert
authors.initials: G
authors.lastname: Lenoir
authors.forename: Li
authors.initials: L
authors.lastname: Mao
authors.forename: François
authors.initials: F
authors.lastname: Janot
mesh_terms: D002294:Carcinoma, Squamous Cell / Q000235:genetics / Q000473:pathology*; D045744:Cell Line, Tumor; D018899:CpG Islands / Q000235:genetics; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics; D019175:DNA Methylation; D006801:Humans; D007012:Hypopharyngeal Neoplasms / Q000235:genetics / Q000473:pathology*; D007822:Laryngeal Neoplasms / Q000235:genetics / Q000473:pathology*; D018895:Microsatellite Repeats / Q000235:genetics; D009367:Neoplasm Staging / Q000379:methods; D010610:Pharyngeal Neoplasms / Q000235:genetics / Q000473:pathology*; D011401:Promoter Regions, Genetic / Q000235:genetics; D011446:Prospective Studies
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16