Background: DNA-dependent protein kinase catalytic subunit (DNA-PK cs ) is known to function in several types of cancer. In this study, we investigated the expression and clinicopathologic significance of DNA-PK cs in laryngeal squamous cell carcinoma (LSCC). Methods: We conducted a retrospective study of 208 patients with advanced-stage LSCC treated at Sun Yat-sen University Cancer Center, Guangzhou, China. We assessed DNA-PK cs and p16INK4 a (p16) status using immunohistochemistry. We examined the association between DNA-PK cs expression and clinicopathologic features and survival outcomes. To evaluate the independent prognostic relevance of DNA-PK cs , we used univariate and multivariate Cox regression models. We estimated overall survival (OS) and distant metastasis-free survival (DMFS) using the Kaplan-Meier method. Results: Immunohistochemical analyses revealed that 163/208 (78.4%) of the LSCC tissue samples exhibited high DNA-PK cs expression. High DNA-PK cs expression was significantly associated with survival outcomes (P = 0.016) and distant metastasis (P = 0.02; chi-squared test). High DNA-PK cs expression was associated with a significantly shorter OS and DMFS than low DNA-PK cs expression (P = 0.029 and 0.033, respectively; log-rank test), and was associated with poor OS in the p16-positive subgroup (P = 0.047). Multivariate analysis identified DNA-PK cs as an independent prognostic indicator of OS and DMFS in all patients (P = 0.039 and 0.037, respectively). Conclusions: Our results suggest that patients with LSCC in whom DNA-PK cs expression is elevated have a higher incidence of distant metastasis and a poorer prognosis. DNA-PK cs may represent a marker of tumor progression in patients with p16-positive LSCC. © Ivyspring International Publisher.