Objective In this study, we expored the enhancing effect of Lupeol on migration and invasion abilities of ST3GalⅢ-silenced MDA-MB-231 cells.  Methods Human breast cancer cell line ST3GalⅢ -silenced MDA-MB-231 was cultured in vitro. The cell adhesion, Transwell and woud healing test were utilized to test the effect of Lupeol on ST3GalⅢ-silenced MDA-MB-231 cells. The expression of matrix metalloproteinase(MMP)-2, -9, phosphatidylinositol 3-kinase/protein kinase B/nuclear factor κB(PI3K/Akt/NF-κB) signaling pathway were examined by Western blotting.  Results No significant influence of the decreased expression of ST3GalⅢ and Lupeol（5μmol/L）on proliferation and apoptosis of MDA-MB-231 cells were found. Lupeol inhibited the migration and invasion of ST3GalⅢ-silenced MDA-MB-231 cells in vitro(P<0.05). Furthermore, the expression of NF-κB p65, p-Akt, Akt, p85, MMP-2 and MMP-9 levels were significantly down-regulated.  Conclusion These observations suggest that Lupeol may inhibit the abilities of migration and invasion of ST3GalⅢ-silenced MDA-MB-231 cells in vitro by inhibiting the protein expression of MMP-2, -9 and effect the signaling pathway of PI3K/Akt/NF-κB.