Objective: To investigate the microRNA-616(miR-616) expression in hepatocellular carcinoma (HCC) and its clinical significance. Methods: The miR-616 expressions in specimens of HCC and adjacent non-tumorous tissue from 80 HCC patients, as well as in normal hepatic cells and different HCC cell lines were determined by real-time PCR. The relations of miR-616 expression with pathologic factors and prognosis of HCC patients were analyzed and the changes in invasion and metastatic abilities in HCC cells after miR-616 overexpression or inhibition were observed. Results: The miR-616 expression in HCC tissue was significantly elevated compared with adjacent non-tumorous liver tissue, and further, in recurrent cases was significantly higher than that in non-recurrent cases and in cases with metastasis was significantly higher than that in cases without metastasis(all P<0.05). The miR-616 expressions were significantly increased in all HCC cell lines compared with normal hepatic cells, which in highly invasive HCC cells were significantly higher than those in lowly invasive HCC cells (all P<0.05). The miR-616 expression was significantly associated with the presence or absence of portal vein tumor thrombus, Edmondson-Steiner grade and TNM stage of HCC patients (all P<0.05), and both overall survival and disease-free survival in HCC patients with high miR-616 expression were significantly lower than those in patients with low miR-616 expression(both P<0.05). The invasive and metastatic abilities in HCC cells were significantly increased after miR-616 overexpression and were significantly reduced after miR-616 inhibition (all P<0.05). Conclusion: MiR-616 expression is increased in HCC, and the high miR-616 expression level is closely associated with poor prognosis.