Abstract
BACKGROUND: Recurrence and metastasis represent the primary causes of therapeutic failure in head and neck squamous cell carcinoma (HNSCC). However, the underlying molecular mechanisms driving these processes remain incompletely elucidated. METHODS: WNK1 (with-no-lysine kinase 1) expression in both normal and HNSCC tissues was analyzed using the TCGA, TIMER2.0 and Clinical Proteomic Tumor Analysis Consortium (CPTAC) datasets, and further validated in HNSCC cell lines by Western blot. Cell Counting Kit-8 (CCK-8), colony formation, Transwell, and orthotopic xenograft assays were conducted to assess the biological role of WNK1 in HNSCC proliferation and metastasis. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) -based quantitative proteomics and post-translational modification profiling were performed to explore the underlying molecular mechanisms. RESULTS: WNK1 overexpression was significantly elevated in HNSCC tissues and cell lines and was correlated with reduced overall survival. WNK1 knockdown suppressed tumor growth and metastasis both in vivo and in vitro. Proteomic and phosphoproteomic profiling of WNK1-driven alterations identified critical signaling pathways closely associated with tumor malignancy. Specifically, WNK1 promotes NF-κB activation through RELA (p65) phosphorylation and nuclear accumulation, resulting in the upregulation of pro-tumorigenic effectors. CONCLUSIONS: Elevated WNK1 drives invasive progression and distant metastasis in HNSCC through NF-κB-dependent transcriptional reprogramming, highlighting its potential as a novel therapeutic target for HNSCC. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12672-025-03637-2.