In this study, we established stable clones of interleukin (IL)-33-overexpressing head and neck squamous cell carcinoma (HNSCC) cells to simulate IL-33-induced autocrine signaling and identify the role of IL-33 expression in more aggressive phenotypes with increased mobility and properties of cancer stemness to provide a potential therapeutic strategy for HNSCC patients, on the basis of targeting the IL-33-enhanced-CXCR4 regulatory circuit. Here, CXCR4 is linked to head and neck squamous cell carcinoma.