Objective To explore the effect of osteopontin (OPN) on pancreatic stellate cell (PSC) and its mechanisms. Methods We transfected PSC with OPN lentiviral overexpression vector (OPN-O/E) and constructed empty vector control cells (control group). After PSCs were treated with OPN-O/E or OPN-O/E in combination with Akt inhibitor LY294002 (10 μmol/L and 50 μmol/L), the proliferation ability and chemotactic activity were detected by CCK-8 and Transwell assays, respectively. The expression levels of α-smooth muscle actin (α-SMA) and related proteins of PI3K/Akt signal pathway were determined by Western blotting. Results Compared with the control group, proliferation ability and chemotactic activity of PCS were significantly increased in the OPN-O/E group, and the expression levels of phosphorylated-PI3K (p-PI3K), phosphorylated-Akt (p-Akt) and α-SMA were also significantly increased (P<0.05 or P<0.01). There were no significant differences in the expression levels of PI3K or Akt between the OPN-O/E and control groups (both P>0.05). Compared with the OPN-O/E group, the expression levels of α-SMA and p-Akt were significantly inhibited in the PCS treated with 10 μmol/L or 50 μmol/L LY294002 (all P<0.01); however, there was no significant difference in the Akt expression. Conclusion OPN can activate PSC through the PI3K/Akt signaling pathway, and the proliferation ability and chemotactic activity of activated PSC are also increased.