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Epstein–Barr virus is an etiologic agent of several malignancies. In this study, we explored the association of Epstein–Barr virus–encoded RNA1 and Epstein–Barr virus latent membrane protein 1 co-expression with osteosarcoma. Epstein–Barr virus–encoded RNA1 expression in tumor cells was quantified using reverse transcriptase polymerase chain reaction and in situ hybridization and Epstein–Barr virus latent membrane protein 1 expression was measured using immunohistochemistry staining. There was a statistically significant association between Epstein–Barr virus latent membrane protein 1 and Epstein–Barr virus–encoded RNA1 co-expression and characteristics of osteosarcoma such as nodal stage (p < 0.04), metastasis (p < 0.04), Ki67 index (p < 0.03), and tumor stage (p < 0.05). Co-expression of Epstein–Barr virus–encoded RNA1 and Epstein–Barr virus latent membrane protein 1 in tumors correlated with advanced osteosarcoma and indicated the aggressiveness of bone sarcoma.