title: No evidence of HPV DNA in esophageal squamous cell carcinoma in a population of Southern Brazil.
abstract: OBJECTIVE
  To investigate the association between human papillomavirus (HPV) and esophageal squamous cell carcinoma (ESCC) in southern Brazil.
  
  
  METHODS
  We studied 189 esophageal samples from 125 patients from three different groups: (1) 102 biopsies from 51 patients with ESCC, with one sample from the tumor and another from normal esophageal mucosa distant from the tumor; (2) 50 esophageal biopsies from 37 patients with a previous diagnosis of head and neck squamous cell carcinoma (HNSCC); and (3) 37 biopsies from esophageal mucosa with normal appearance from 37 dyspeptic patients, not exposed to smoking or alcohol consumption. Nested-polymerase chain reaction (PCR) with the MY09/11 and GP5/6 L1 primers was used to detect HPV L1 in samples fixed in formalin and stored in paraffin blocks. All PCR reactions were performed with a positive control (cervicovaginal samples), with a negative control (Human Genomic DNA) and with a blank reaction containing all reagents except DNA. We took extreme care to prevent DNA contamination in sample collection, processing, and testing.
  
  
  RESULTS
  The histological biopsies confirmed the diagnosis of ESCC in 52 samples (51 from ESCC group and 1 from the HNSCC group) and classified as well differentiated (12/52, 23.1%), moderately differentiated (27/52, 51.9%) or poorly differentiated (7/52, 13.5%). One hundred twenty-eight esophageal biopsies were considered normal (51 from the ESCC group, 42 from the HNSCC group and 35 from dyspeptic patients). Nine had esophagitis (7 from the HNSCC and 2 from dyspeptic patients). Of a total of 189 samples, only 6 samples had insufficient material for PCR analysis: 1 from mucosa distant from the tumor in a patient with ESCC, 3 from patients with HNSCC and 2 from patients without cancer. In 183 samples (96.8%) GAPDH, G3PDH and/or β-globin were amplified, thus indicating the adequacy of the DNA in those samples. HPV DNA was negative in all the 183 samples tested: 52 with ESCC, 9 with esophagitis and 122 with normal esophageal mucosa.
  
  
  CONCLUSIONS
  There was no evidence of HPV infection in different ESCC from southern Brazil.
authors.affiliation: Luís Carlos Moreira Antunes, João Carlos Prolla, Antonio de Barros Lopes, Renato Borges Fagundes, Programa de Pós-Graduação: Ciências em Gastroenterologia e Hepatologia - Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul 90035-003, Brasil.
authors.forename: Luís Carlos Moreira
authors.initials: LC
authors.lastname: Antunes
authors.forename: João Carlos
authors.initials: JC
authors.lastname: Prolla
authors.forename: Antonio
authors.initials: A
authors.lastname: de Barros Lopes
authors.forename: Marta Pires
authors.initials: MP
authors.lastname: da Rocha
authors.forename: Renato Borges
authors.initials: RB
authors.lastname: Fagundes
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001706:Biopsy; D001938:Brazil / Q000453:epidemiology; D002294:Carcinoma, Squamous Cell / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D004279:DNA, Viral / Q000032:analysis*; D004938:Esophageal Neoplasms / Q000453:epidemiology / Q000473:pathology / Q000821:virology*; D000077277:Esophageal Squamous Cell Carcinoma; D005260:Female; D061809:Human Papillomavirus DNA Tests; D006801:Humans; D008297:Male; D008875:Middle Aged; D027383:Papillomaviridae / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis / Q000453:epidemiology / Q000821:virology*; D016133:Polymerase Chain Reaction; D011237:Predictive Value of Tests; D012307:Risk Factors; D055815:Young Adult
chemical_list: D004279:DNA, Viral
keywords: Esophageal cancer; Esophageal squamous cell carcinoma; Head and neck cancer; Human papillomavirus; Nested-polymerase chain reaction; Polymerase chain reactions