No role for human papillomavirus in esophageal squamous cell carcinoma in China.
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.
Koshiol
Jill
J
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
Wei
Wen-Qiang
WQ
Kreimer
Aimee R
AR
Chen
Wen
W
Gravitt
Patti
P
Ren
Jian-Song
JS
Abnet
Christian C
CC
Wang
Jian-Bing
JB
Kamangar
Farin
F
Lin
Dong-Mei
DM
von Knebel-Doeberitz
Magnus
M
Zhang
Yu
Y
Viscidi
Raphael
R
Wang
Guo-Qing
GQ
Gillison
Maura L
ML
Roth
Mark J
MJ
Dong
Zhi-Wei
ZW
Kim
Esther
E
Taylor
Philip R
PR
Qiao
You-Lin
YL
Dawsey
Sanford M
SM
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*
D004279:DNA, Viral