PFKP binding AMOTL1 promotes tumor aerobic glycolysis and epithelial-mesenchymal transition by modulating Hippo pathway in head and neck cancer.

BACKGROUND: Aerobic glycolysis drives cancer progression through phosphofructokinase (PFK)-mediated regulation. The contribution of platelet-type PFK (PFKP) to head and neck squamous cell carcinoma (HNSCC) pathogenesis remains undefined. METHODS: Bioinformatic screening of 548 TCGA-HNSCC cases identified glycolysis-related prognostic genes via Cox regression, followed by experimental validation using quantitative polymerase chain reaction (qPCR) and Kaplan-Meier survival analysis in 51 clinical HNSCC tissue pairs in our center. In vitro functional assays (CCK8, colony formation, migration assays and metabolic analysis experiments), co-immunoprecipitation, ubiquitination analysis, and immunofluorescence were performed in HNSCC cell lines. PFKP-AMOTL1 interaction was validated by protein-binding studies. In vivo tumor growth was assessed using nude mice models. Clinical correlation analysis utilized HNSCC patient cohorts. Statistical significance was determined by Student's t-test and ANOVA. RESULTS: PFKP emerged as the pivotal prognostic biomarker for HNSCC, demonstrating significant upregulation at mRNA/protein levels in tumors versus normal tissues (TCGA/clinical cohorts) and correlating with reduced overall and disease-specific survival. In vitro, PFKP overexpression enhanced aerobic glycolysis and epithelial-mesenchymal transition (EMT) in HNSCC cells. Mechanistically, PFKP directly bound AMOTL1, inhibiting its ubiquitin-mediated degradation. PFKP-driven glycolysis and EMT were AMOTL1-dependent. Furthermore, PFKP promoted YAP nuclear translocation via AMOTL1, suppressing Hippo pathway activity and amplifying glycolytic flux. In vivo, elevated PFKP accelerated tumor progression and glycolytic metabolism through AMOTL1/YAP/Hippo signaling. CONCLUSIONS: PFKP remodels tumor metabolism and drives EMT in HNSCC via the AMOTL1/YAP/Hippo axis, highlighting its mechanistic and prognostic significance, and suggesting its potential as a future therapeutic target.