Abstract
Human papillomavirus type 16 (HPV16) is the most common type of infection in head and neck squamous cell carcinoma (HNSCC).The HPV16-encoded miRNA, HPV16-miR-H1, has been validated in cervical cancer tissues and cells, but its role in the pathogenesis of HNSCC remains unclear. The expression level of HPV16-miR-H1 in HNSCC cells was detected, and its mechanism of action was analyzed using bioinformatics tools and RNA sequencing. Overexpression and knockdown of HPV16-miR-H1 were performed to validate its regulatory effects on the PTEN/AKT axis and its impact on cell proliferation, migration, the cell cycle, and glycolysis. HPV16-miR-H1 was highly expressed in HPV16-positive HNSCC cells. Bioinformatics prediction and gene enrichment analysis indicated that HPV16-miR-H1 regulates PTEN transcription and the glycolysis process. Dual-luciferase reporter assays further confirmed that HPV16-miR-H1 targets the 3’-UTR region of PTEN. Functional experiments demonstrated that HPV16-miR-H1 affects HNSCC cell proliferation, migration, the cell cycle, and glycolysis. The expression of HPV16-miR-H1 in HNSCC cells was validated for the first time. Further studies revealed that it regulates the PTEN/AKT axis by targeting the 3’UTR region of PTEN, thereby enhancing the proliferation, migration, and glycolytic processes in HNSCC cells. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1038/s41598-025-31764-x.