A mass spectrometry-based proteomics study carried out on 20 oral squamous cell carcinoma (OSCC) samples identified 26 differentially expressed proteins between high-risk human papillomavirus DNA (HR-HPV DNA) positive (+) and negative (-) tumors. Among these proteins, S100-A8 was upregulated in HR-HPV + cases and presented the most relevant correlation with survival rates. Objective: Validate the expression of S100-A8 in an expanded number of OSCC and correlate it with clinicopathological features and survival outcomes. Study Design: Immunofluorescence for S100-A8 was performed on a 91 OSCC tissue microarray. Additionally, The Cancer Genome Atlas data set of 338 OSCC samples was analyzed for S100-A8 mRNA levels. Results: Higher expression of S100-A8 protein and mRNA levels in HR-HPV DNA + tumors were confirmed. The up-regulated S100-A8 was correlated with T3/T4 tumors, advanced clinical stage and positive surgical margins. Furthermore, a high level of S100-A8 was associated with decreased disease-free survival, cancer specific, and overall survival. Conclusions: S100-A8 may play a role in HR-HPV DNA-related OSCC onset and progression. Acknowledgments: This work was supported by FAPESP grants: 2016/03248-1 and 2017/16347-0. We acknowledge the Mass Spectrometry Laboratory at Brazilian Biosciences National Laboratory, CNPEM, Campinas, Brazil for their support with the mass spectrometry analysis.