title: No role for human papillomavirus in esophageal squamous cell carcinoma in China.
abstract: Certain regions of China have high rates of esophageal squamous cell carcinoma (ESCC). Previous studies of human papillomavirus (HPV), a proposed causal factor, have produced highly variable results. We attempted to evaluate HPV and ESCC more definitively using extreme care to prevent DNA contamination. We collected tissue and serum in China from 272 histopathologically-confirmed ESCC cases with rigorous attention to good molecular biology technique. We tested for HPV DNA in fresh-frozen tumor tissue using PCR with PGMY L1 consensus primers and HPV16 and 18 type-specific E6 and E7 primers, and in formalin-fixed paraffin-embedded tumor tissue using SPF(10) L1 primers. In HPV-positive cases, we evaluated p16(INK4a) overexpression and HPV E6/E7 seropositivity as evidence of carcinogenic HPV activity. beta-globin, and thus DNA, was adequate in 98.2% of the frozen tumor tissues (267/272). Of these, 99.6% (95% confidence interval (CI) = 97.9-100.0%) were negative for HPV DNA by PGMY, and 100% (95% CI = 98.6-100%) were negative by HPV16/18 E6/E7 PCR. In the corresponding formalin-fixed paraffin-embedded tumor specimens, 99.3% (95% CI = 97.3-99.9%) were HPV negative by SPF(10). By PGMY, 1 case tested weakly positive for HPV89, a noncancer causing HPV type. By SPF(10), 2 cases tested weakly positive: 1 for HPV16 and 1 for HPV31. No HPV DNA-positive case had evidence of HPV oncogene activity as measured by p16(INK4a) overexpression or E6/E7 seropositivity. This study provides the most definitive evidence to date that HPV is not involved in ESCC carcinogenesis in China. HPV DNA contamination cannot be ruled out as an explanation for high HPV prevalence in ESCC tissue studies with less stringent tissue procurement and processing protocols.
authors.affiliation: Division of Cancer Epidemiology and Genetics, Department of Health and Human Services, National Cancer Institute, National Institutes of Health, Bethesda, MD 20852-7248, USA. koshiolj@mail.nih.gov
authors.forename: Jill
authors.initials: J
authors.lastname: Koshiol
authors.forename: Wen-Qiang
authors.initials: WQ
authors.lastname: Wei
authors.forename: Aimee R
authors.initials: AR
authors.lastname: Kreimer
authors.forename: Wen
authors.initials: W
authors.lastname: Chen
authors.forename: Patti
authors.initials: P
authors.lastname: Gravitt
authors.forename: Jian-Song
authors.initials: JS
authors.lastname: Ren
authors.forename: Christian C
authors.initials: CC
authors.lastname: Abnet
authors.forename: Jian-Bing
authors.initials: JB
authors.lastname: Wang
authors.forename: Farin
authors.initials: F
authors.lastname: Kamangar
authors.forename: Dong-Mei
authors.initials: DM
authors.lastname: Lin
authors.forename: Magnus
authors.initials: M
authors.lastname: von Knebel-Doeberitz
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
authors.forename: Raphael
authors.initials: R
authors.lastname: Viscidi
authors.forename: Guo-Qing
authors.initials: GQ
authors.lastname: Wang
authors.forename: Maura L
authors.initials: ML
authors.lastname: Gillison
authors.forename: Mark J
authors.initials: MJ
authors.lastname: Roth
authors.forename: Zhi-Wei
authors.initials: ZW
authors.lastname: Dong
authors.forename: Esther
authors.initials: E
authors.lastname: Kim
authors.forename: Philip R
authors.initials: PR
authors.lastname: Taylor
authors.forename: You-Lin
authors.initials: YL
authors.lastname: Qiao
authors.forename: Sanford M
authors.initials: SM
authors.lastname: Dawsey
mesh_terms: D000368:Aged; D002294:Carcinoma, Squamous Cell / Q000473:pathology / Q000821:virology*; D002681:China; D004279:DNA, Viral / Q000235:genetics; D004938:Esophageal Neoplasms / Q000473:pathology / Q000821:virology*; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D027383:Papillomaviridae / Q000235:genetics / Q000472:pathogenicity*
chemical_list: D004279:DNA, Viral