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 SPF10 L1 primers. In HPV-positive cases, we evaluated p16INK4a overexpression and HPV E6/E7 seropositivity as evidence of carcinogenic HPV activity. β-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 SPF10. By PGMY, 1 case tested weakly positive for HPV89, a noncancer causing HPV type. By SPF10, 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 p16INK4a 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.

Prevalence of Human Papillomavirus 16 in Esophageal Cancer Among the Chinese Population: a Systematic Review and Meta-analysis

“…Furthermore, eight studies were excluded because of the limited number. In total, we included 26 eligible studies in the meta-analysis (Cao et al, 2005;Yang et al, 2005;Dai et al, 2007;He et al, 2007;Shuyama et al, 2007;Zhou et al, 2007;Chen et al, 2008;Li et al, 2008;Liu et al, 2008;Lu et al, 2008;Zhao et al, 2009;Liu et al, 2010;Koshiol et al, 2010;Wang et al, 2010;Ayshamgul et al, 2011;Han et al, 2011;Zhang et al, 2011;Guo et al, 2012;Hu et al, 2012;Qu et al, 2012;Liu et al, 2013;Zhang et al, 2013;Cui et al, 2014;Liu et al, 2014).…”

Revisit of Field Cancerization in Squamous Cell Carcinoma of Upper Aerodigestive Tract: Better Risk Assessment with Epigenetic Markers

“…However, judgment-based merely on the overall exposure to ABC was insufficient because the false-positive rate (55%) was high. Both of the infectious pathogens H. pylori and human papillomavirus were not associated with SCC risk (37,38). Although several risk alleles have been previously identified (10-13), they served as effect modifiers rather than direct causes.…”

Human Papillomavirus Infection and Esophageal Squamous Cell Carcinoma: A Case–Control Study

“…However, the presence of HPV DNA in esophageal carcinomas has been a highly controversial issue (16,20,22,32). Differences in sampling methods, demographic and ethnic factors, disease status, and sensitivity of detection methods have been suggested to be potential causes of the discrepancies in findings (16).…”

“…We have frequently detected HPV DNA in esophageal squamous cell carcinoma (ESCC) tissues collected from patients in Anyang (19), a high-incidence area for ESCC in China, whereas other studies failed to produce comparable results in samples collected from same region and concluded there was no association between HPV infection and ESCC (20)(21)(22). Our previous study (refer to the Supplementary data) and a study from other investigators have observed that HPV viral load in ESCC is much lower than in cervical cancer tissues (23).…”