Objective To investigate the effect of osteopontin (OPN) interference on the Warburg effect of hepatoma cells and the related molecular mechanism. Methods Small interfere RNA (siRNA) targeting OPN (si-OPN) and negative control (si-NC) were transfected into HepG2 and SK-HEP-1 cells. 2-NBDG fluorescent probe was used to detect the glucose uptake capacity of hepatoma, and the lactate detection kit was used to evaluate the lactate production in HepG2 and SK-HEP-1 cells. The expression of glycolysis-related genes, suchas GLUT1, HK2 and LDHA was assessed by RT-qPCR and western blot assays. After silencing the expression of LGALS3BP in HepG2 and SK-HEP-1 cells, Western blot was used to detect the expression of LGALS3BP, GLUT1, HK2 and LDHA. The glucose uptake and lactate production were also evaluated in LGALS3BP silenced HepG2 and SK-HEP-1 cells. Results Compared with the control group and si-NC group, the levels of glucose uptake and lactate production were significantly reduced in HepG2 and SK-HEP-1 cells interfe