Metastasis suppressor 1 (MTSS1) is a potential metastasis suppressor gene involved in the regulation of cytoskeleton dynamics and subsequently in cell motility. MTSS1 expression is frequently reduced in a variety of cancer cells and tissues and this loss may account for increased invasive traits in cancer cells. The present study aimed to assess the role of MTSS1 in epithelial ovarian cancer (EOC) cells. Expression of MTSS1 in human ovarian cells was assessed at both the mRNA and protein levels using reverse transcription-PCR (RT-PCR) and immunohistochemistry, respectively. Full-length MTSS1 cDNA expression vector was used to generate MTSS1 overexpressing cells. The effect of MTSS1 overexpression on cellular functions was examined in EOC cells using a variety of in vitro assays. MTSS1 expression was observed both in ovarian cancer tissues and EOC cells. Over-expression of MTSS1 protein reduced the growth, invasion, adhesion and migration of EOC cell lines in vitro. The present study revealed that MTSS1 plays an essential inhibitory role in the development and progression of ovarian cancers. MTSS1 overexpression is intimately related to migration and metastasis, suggesting that MTSS1 is a potential prognostic marker and therapeutic molecular target in human ovarian cancer.