BACKGROUND: Circular RNAs (circRNA) are involved in the progression of cancers, and previous study showed that hsa_circ_0001649 expression is down-regulated in hepatocellular carcinoma (HCC). OBJECTIVE: To explore whether hsa_circ_0001649 is a prognostic biomarker for HCC and to investigate the biological functions of hsa_circ_0001649 in HCC. METHODS: Hsa_circ_0001649 expression was measured in 77 pairs of HCC and adjacent no-tumor tissues by quantitative Real-Time polymerase chain reaction. Kaplan-Meier curve and Cox regression were used to analyze its prognostic significance for HCC patients. In addition, the hsa_circ_0001649 was over-expressed using a circRNA-forming plasmid in HCC cells, and the biological function of hsa_circ_0001649 was investigated in vitro. RESULTS: We verified that hsa_circ_0001649 was down-regulated in HCC tissues compared with adjacent non-tumor tissues. In addition, low hsa_circ_0001649 expression was associated with the poor overall survival of HCC patients, and Cox multivariate analysis showed that hsa_circ_0001649 is a novel independent prognostic factor for HCC patients. Furthermore, the in vitro experiments demonstrated that over-expressed hsa_circ_0001649 inhibits the proliferation, migration, and invasion and promotes the apoptosis of HCC cells. CONCLUSIONS: Hsa_circ_0001649 could act as a novel prognostic biomarker for HCC patients. In addition, hsa_circ_0001649 might be a potential therapeutic target for HCC.