Assessment of Immunohistochemistry for p16INK4 and High-Risk HPV DNA by In Situ Hybridization in Esophageal Squamous Cell Carcinoma

Abstract

The role of high-risk human papillomavirus (HPV) in the pathogenesis of esophageal squamous cell carcinoma (ESCC) remains unclear. p16(INK4) is used as a surrogate marker to detect HPV-related tumors but has had discrepant results in ESCC. In this study, 32 cases of ESCC were examined to determine the relationship between p16(INK4) expression and high-risk HPV. All the tumors were stained by immunohistochemistry for p16(INK4). Tumors having p16(INK4) nuclear and/or nuclear and cytoplasmic expression were considered positive. Tumors positive for p16(INK4) expression were tested for high-risk HPV by in situ hybridization (ISH). In all, 20 cases of ESCC (63%) showed only cytoplasmic staining for p16(INK4), and 11 cases (34%) showed both cytoplasmic and nuclear staining for p16(INK4); 4 cases (13%) showed no staining for p16(INK4). None of the p16(INK4) -positive cases were positive for high-risk HPV by ISH. These results indicate that p16(INK4) expression in ESCC does not correlate with the presence of high-risk HPV DNA by ISH. High-risk HPV does not seem to play a major role in the carcinogenesis of ESCC in low-risk areas.

Role of human papillomaviruses in esophageal squamous cell carcinoma

“…In general, studies conducted in high‐risk regions for ESCC incidence, have reported higher rates of HPV in ESCC specimens than studies that are carried out in low‐risk regions. Many of the most recent studies continue to support this trend . However, there have been some exceptions to this pattern.…”

Mucosal alpha‐papillomaviruses are not associated with esophageal squamous cell carcinomas: Lack of mechanistic evidence from South Africa, China and Iran and from a world‐wide meta‐analysis

“…37 And while 14 studies have reported on expression of p16 INK4a protein in ESCC tissues, they used varying cut-offs to define p16 INK4a positivity and reported contradicting results. 15,34,[38][39][40][41][42][43][44][45][46][47][48][49] As a result of inconsistent HPV DNA data and very limited direct functional evidence provided in the last 30 years of research, the worldwide expert panel of the 100 th IARC Monograph concluded that the evidence for a causal association between HPV and ESCC remains inconclusive. 4 In addition to HPV functional markers that can be assessed in tumor tissues, antibodies to HPV early proteins, especially E6 and E7, have been demonstrated to be markers for HPV-driven SCC of the cervix, 50 penis 51 and oropharynx.…”

“…Fourteen previous studies which reported data on both HPV DNA testing and expression of p16 INK4a as an HPV functional marker were identified for possible inclusion in the meta-analysis. 15,34,[38][39][40][41][42][43][44][45][46][47][48][49] Of these 14 studies, 5 were excluded for the following reasons: Antonsson et al 38 and Koshiol et al 15 HPV DNA1 cases in their ESCC series and hence the study provides no information about relative probabilities of HPV DNA1 and p16 INK4a 1 39 ; and Vaiphei et al 49 did not report specific frequencies of p16 INK4a 1 tumors. The remaining 9 studies, and our own findings (i.e., 10 studies in total), were included in the meta-analysis.…”

“…Scarce evidence has been reported on HPV viral load or expression of viral transcripts . And while 14 studies have reported on expression of p16 INK4a protein in ESCC tissues, they used varying cut‐offs to define p16 INK4a positivity and reported contradicting results . As a result of inconsistent HPV DNA data and very limited direct functional evidence provided in the last 30 years of research, the worldwide expert panel of the 100 th IARC Monograph concluded that the evidence for a causal association between HPV and ESCC remains inconclusive…”

Correlation among 16 biological factors [p53, p21waf1, MIB-1 (Ki-67), p16INK4A, cyclin D1, E-cadherin, Bcl-2, TNF-α, NF-κB, TGF-β, MMP-7, COX-2, EGFR, HER2/neu, ER, and HIF-1α] and clinical outcomes following curative chemoradiation therapy in 10 patients with esophageal squamous cell carcinoma

“…In previous studies, it was considered that the expression levels of p53 (1–3), p21 waf1 (4), molecular immunology borstel-1 (MIB-1) (2,5), p16 INK4A (6), cyclin D1 (2,7,8), E-cadherin (8), Bcl-2 (1), tumor necrosis factor (TNF)-α (9), nuclear factor (NF)-κB (10), transforming growth factor (TGF)-β (11), matrix metalloproteinase (MMP)-7 (12), cyclooxygenase (COX)-2 (13,14), epidermal growth factor receptor (EGFR) (15) and hypoxia-inducible factor (HIF)-1α (16) may be used as prognostic factors for ESCC. However, these studies included patients treated with surgery or neoadjuvant chemoradiation therapy (CRT).…”