Simple SummaryThe role of human papillomavirus (HPV) in the survival of patients with head and neck squamous cell carcinoma (HNSCC) is an important topic. The recognition of additional markers could play a significant role in survival prognosis. Our study aimed to assess the roles of different molecular and immunohistochemical factors in the survival of patients with HNSCC. We analyzed 106 HNSCC samples and confirmed the roles of HPV DNA and p16, p53, and HPV16 E6 and E7 proteins in different subgroups of HNSCC. In addition to p16, the immunohistochemical overexpression of HPV16 E6 protein should be used for patient survival prognosis.AbstractThe role of HPV in the survival prognosis of patients with head and neck squamous cell carcinoma, especially patients with laryngeal squamous cell carcinoma (LSCC) and hypopharyngeal squamous cell carcinoma (HPSCC), is still somewhat ambiguous. The present study aimed to explore the significance of tumor suppressor proteins and HPV16 E6 and E7 oncoproteins in the assessment of survival in patients with oropharyngeal squamous cell carcinoma (OPSCC), LSCC, and HPSCC associated with high-risk (HR-) and low-risk (LR-) HPV infections. By utilizing molecular and immunohistochemical investigations of HNSCC samples and patient data, univariate and multivariate survival analyses were conducted. The presence of HPV DNA (LR- and HR-HPV) was associated with a better 5-year OS and DSS for OPSCC and LSCC. The IHC overexpression of HPV16 E6 protein and p16 protein was associated with better survival in the univariate (for OPSCC) and multivariate (OPSCC and HPSCC) survival analyses. The overexpression of p53 was associated with better survival in OPSCC. HPV infection plays a significant role in the tumorigenesis of HNSCC, and the immunohistochemical assessment of HPV16 E6 protein expression should be interpreted as a useful prognostic marker for OPSCC and HPSCC.