It has been reported that MicroRNAs (miRNAs) play pivotal roles in the occurrence and progression of a variety of cancers. As reported, miR-4295 promotes cell growth and metastasis in a lot of cancers. Nonetheless, the role and molecular mechanism of miR-4295 in HNSCC still remain unknown. In this study, we discovered miR-4295 expression was significantly upregulated in HNSCC tissues and cell lines, which is also associated with the overall survival of patients. Additionally, suppression of miR-4295 significantly inhibited cell proliferation, migration and EMT process in HNSCC. Through Targetscan website, it was predicted that NPTX1 might be a direct target gene of miR-4295. Then, we verified that NPTX1 could directly interact with miR-4295 via luciferase reporter and RNA assays. What's more, we discovered that there was a significantly negative correlation between NPTX1 and miR-4295 expression. It was indicated by further investigation that the effect of miR-4295 suppression on cell proliferation, migration and EMT process in HNSCC can be restored by knockdown of NPTX1 at the same time. Our results suggested that miR-4295 promoted the progression of HNSCC via regulating NPTX1 expression and miR-4295/NPTX1 axis, which may be a new therapeutic strategy for HNSCC.