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Head and neck squamous cell carcinoma (HNSCC) is one of the most common types of human cancer. Typically HNSCC cells show persistent invasion that frequently leads to local recurrence and distant lymphatic metastasis. However, molecular mechanisms associated with invasion and metastasis of HNSCC remain poorly understood. Here we identified Periostin as an invasion promoting factor in HNSCC by comparing the gene expression profiles between parent HNSCC cells and a highly invasive clone. Indeed, Periostin overexpression promoted the invasion and anchorage independent growth both in vitro and in vivo in HNSCC cells. Moreover, Periostin overexpressing cells spontaneously metastasized to cervical lymph nodes and to the lung through their aggressive invasiveness in an orthotopic mouse model of HNSCC. Interestingly, Periostin was highly expressed in HNSCCs in comparison with normal tissues, and the level of Periostin expression was well correlated with the invasiveness of HNSCC cases. In summary, these findings suggest that Periostin plays an important role for invasion and anchorage independent growth in the metastatic process of HNSCC.