The purpose of this study was to investigate significance of HPV16 DNA/p16 for survival of patients with laryngeal squamous cell carcinoma (LSCC). HPV16 DNA and p16 status in 812 LSCC patients were determined. The survival was performed using Kaplan-Meier estimates and Cox model. Compared with HPV negativity, adjusted HRs for HPV16 positivity were 0.57, 95% CI, 0.38 to 0.87; 0.60, 0.40 to 0.88; and 0.65, 0.46 to 0.94 for disease-specific survival (DSS), recurrence-free survival (RFS), and overall survival (OS), respectively. Additionally, for p16 expression, aHRs were 0.68, 95% CI, 0.48 to 0.96; 0.72, 0.52 to 0.98; and 0.73, 0.54 to 0.99 for DSS, RFS, and OS, respectively. Finally, for combined analysis, patients with both HPV16-positivity/p16-positivity had much better prognosis than those with either negativity. Such above survivals are more significantly better in never smokers. Our findings suggest that HPV16/p16 may affect survival outcomes of LSCC patients, particularly in never smokers. © 2019 Wiley Periodicals, Inc.