Background: CircRNAs involved in the development of many diseases have been recently identified. However, the possible role of circRNAs in human hepatocellular carcinoma (HCC) remains to be investigated. Objective: This study aimed to identify the expression of hsa_circ_0085616 in different HCC cell lines and its function in HCC tumorigenesis. Methods: The expression of hsa_circ_0085616 was first measured in 68 pairs of HCC tissues and matched normal adjacent tissues. Further analysis was performed in different HCC cell lines and human normal hepatic cell lines. Moreover, we down- or upregulated its expression by cell transfection in vitro. Cell proliferation, migration, invasion and apoptosis were measured, and proliferation-related signaling pathway proteins were also analyzed. Results: We found that hsa_circ_0085616 was highly expressed in all HCC cell lines compared to the normal liver cell line. Upregulation or downregulation of hsa_circ_0085616 expression could strengthen or weaken the proliferative ability of HCC cells in vitro. Moreover, the protein levels of beta-catenin, p-ERK, and p-AKT, which play important roles in the typical proliferation pathway, were also affected by the expression of hsa_circ_0085616. Conclusion: Our study indicated that hsa_circ_0085616 might be a potential biomarker and a new therapeutic target for HCC.