In the current study, miR-328-3p can increase the phosphorylation protein level of mTOR, P70S6K, S6RP and 4E-BP1, and overexpression of the target gene H2AFX reduced the protein levels of p-mTOR, p-p70S6K and p-4E-BP1, suggesting that miR-328-3p/H2AFX axis could activate the mTOR pathway in HNSCC. This evidence concerns the gene RPS6KB1 and head and neck squamous cell carcinoma.