Introduction and objective. Epstein-Barr virus (EBV) is associated with cancers of the head and neck, including oropharyngeal cancer, which is increasing in incidence, and biomarker studies have potential in diagnostics and therapy. One of the most commonly deregulated microRNAs in cancers is miR-21-5p. It has been implicated in neoplastic transformation related to EBV infection in several investigations. The aim of this study was to determine the level of miR-21-5p in the serum of EBV (+) and EBV (-) oropharyngeal cancer patients. Material and methods. The study was carried out on 78 patients with confirmed OPSCC. Statistical analysis was used to investigate the relationship between clinical and demographic characteristics of patients. Enzyme immunoassays were used to determine the levels of miRNA, TLR9 and MMPs and cytokines. Statistical analysis was used to determine the relationship between miR21-5p and TLR9, MMP3, MMP9 levels, and the cytokines studied. Results. Significantly higher values of all tested parameters for miR-21-5p levels and grading as well as TN stage were found in the EBV (+) group. There was no statistically significant correlation between the miR-21-5p level and the levels of TNFα, VEGF, and TGFβ. Positive correlations were shown between miR-21-5p and IL-10, MMP-3 and -9. There was a negative correlation between the level of miR-21-5p and TLR9. Conclusions. The present study showed that in EBV (+) patients the level of miR-21-5p in the serum was significantly higher than in EBV (-) patients. Our study results could influence future strategies for the diagnosis, prevention and treatment of oropharyngeal cancers.