Head and neck squamous cell carcinoma (HNSCC) is a globally prevalent malignancy associated with high incidence and mortality rates, posing a significant threat to patient survival and quality of life (Bray et al., 2024). Despite ongoing advancements in multimodal treatments (including radical surgery, intensity-modulated radiotherapy, and systemic chemotherapy) the 5-year survival rate remains suboptimal at approximately 45 %–50 % (Chi et al., 2022, Bhatia and Burtness, 2023). High rates of postoperative recurrence and distant metastasis continue to limit long-term survival outcomes (Mody et al., 2021). Although targeted therapies and immune checkpoint inhibitors have introduced novel therapeutic options for patients with advanced disease, the lack of personalized treatment strategies and the emergence of drug resistance remain substantial clinical challenges (Lamenza et al., 2025). Therefore, elucidating the molecular mechanisms underlying HNSCC and identifying clinically relevant biomarkers is essential for enhancing diagnostic and therapeutic strategies.

The AKT signaling pathway plays a pivotal role in cancer progression. Dysregulation of its components, including PI3K hyperactivation, PTEN loss, and activating mutations in AKT, drives tumorigenesis and contributes to therapy resistance (Adon et al., 2025). Studies have shown that HPV oncogenes E6/E7 can disrupt the regulation of the EGFR/PI3K/AKT pathway and promote PIK3CA mutations, resulting in widespread pathway dysregulation in both HPV-positive and HPV-negative HNSCC patients (Janecka-Widła et al., 2021). Although this pathway is considered a promising therapeutic target, its upstream regulatory mechanisms remain inadequately characterized.

Twinfilin actin-binding protein 1 (TWF1) is a highly conserved modulator of cytoskeletal dynamics (Shekhar et al., 2021). It contains tandem actin depolymerization factor homology domains, enabling dual regulation of actin turnover by binding actin monomers and interacting with capping proteins to stabilize filaments (Mwangangi et al., 2021, Courtemanche and Henty-Ridilla, 2024). Recent studies have highlighted its oncogenic functions across several malignancies. For example, in lung adenocarcinoma, TWF1 overexpression is associated with poor prognosis and promotes tumor progression through MMP1 activation (Zhai et al., 2023). In breast cancer, TWF1 interacts with MCTS1 to regulate the translation of Cyclin D1 and c-Myc, thereby enhancing proliferation (Tian et al., 2020). In clear cell renal cell carcinoma, TWF1 is a direct target of miR-30a/c-5p and facilitates epithelial–mesenchymal transition, invasiveness, and metastasis (Outeiro-Pinho et al., 2022). These findings suggest that TWF1 contributes to malignant transformation in diverse tumor types.

However, despite these observations, the expression pattern, clinical significance, and mechanistic role of TWF1 in HNSCC remain largely unexplored. Given the central role of the AKT pathway in HNSCC, we hypothesized that TWF1 may drive tumor progression by regulating AKT phosphorylation. To address this, we conducted a systematic investigation integrating TCGA pan-cancer analysis, clinical tissue validation, and in vitro functional assays to clarify the oncogenic role of TWF1 in HNSCC and its potential as a therapeutic target.