Abstract: Objective To investigate the effect and mechanism of osteopontin (OPN) on hepatoma cell migration through galectin 3 binding protein (LGALS3BP). Methods Human hepatoma cell lines SMMC-7721, SMMC-P (stably transfected with empty eukaryotic expression vectors) and SMMC-OPN (stably transfected with OPN gene) were cultured. The mRNA expression levels of OPN and LGALS3BP were detected by qRT-PCR. Western blot assay was used to analyze the relative expressions of OPN and LGALS3BP. The protein expression level of PI3K/AKT pathway, including p-PI3K, PI3K, p-AKT and AKT were detected by western blot. Wound healing assay was performed to explore the cell migration ability. After transfection with LGALS3BP-targeting small interfering RNA (si-LGALS3BP) and negative control small RNA (si-NC) into SMMC-OPN cells respectively, cell migration ability and the relative expression of PI3K/AKT pathway related proteins were detected. Results Compared with SMMC-7721 and SMMC-P, the migration ability of SMMC-OPN cells was significantly reinforced, and the expression of LGALS3BP was obviously up-regulated at both mRNA and protein. Meanwhile, the relative expression quantity of p-PI3K/PI3K and p-AKT/AKT proteins were significantly increased. Wound healing assay showed that the si-LGALS3BP could obviously suppress the migration ability of SMMC-OPN cells. Furthermore, the relative expression of p-AKT/AKT and p-PI3K/PI3K proteins in SMMC-OPN cells were significantly decreased after transfection with si-LGALS3BP. Conclusion OPN could activate PI3K/AKT pathway through up-regulating LGALS3BP expression in promoting hepatoma cells migration.