BackgroundLymph nodes metastasis is the main metastasis mode of tongue squamous cell carcinoma (TSCC). Ras related GTP binding protein B (RalB) have been recently described that it was involved in tumor growth and metastasis, but the effect in TSCC is still ill-defined.ObjectiveThis study provides insights into the role of RALB as a prognostic factor in head and neck squamous cell carcinoma (HNSCC) and demonstrates its involvement in promoting lymph node metastasis in TSCC.MethodsFirstly, the expression level of RALB and the relationship with clinical features were examined. Subsequently, RALB knockdown Cal-27 cells orthotopic xenotransplantation in the tongue of BALB/c nude mice were established. Finally, using Connectivity Map (CMAP) database to find possible drugs.ResultsFirstly, RALB could not only predict the cancer patients' prognosis and survival and but also act as a potential prognostic factor, particularly in HNSCC by pan-cancer bioinformatics analysis. In addition, we found that RalB promoted tumor growth and lymph node metastasis. Finally, we identified Tirabrutinib (ONO-4059) targeting RalB with good binding properties.ConclusionsRalB act as a prognostic gene in HNSCC, and promote lymph node metastasis in early stage of TSCC.