● Wnt/β-catenin signaling pathway is activated in CSCs cell lines and β-catenin overexpression drives the acquisition of CSCs properties as self-renewal, stem cell marker expression, including Oct4, and chemoresistance● β-catenin directly regulates Oct4 transcription in CSCs and Oct4 overexpression abrogates the inhibition of stemness caused by β-catenin knockdown in CSCs● Wnt/β-catenin axis mediates the self-renewal of CSCs in HNSCC● Novel therapeutic strategies for targeting CSCs in HNSCC may focus on the blockade of the Wnt/β-catenin signaling pathway. This evidence concerns the gene POU5F1 and head and neck squamous cell carcinoma.