Objective To investigate the effect of vitamin K deficiency or protein-induced protein Ⅱ (PIVKA-Ⅱ) on the proliferation and metastasis of hepatoma cells and to explore its possible mechanism. Methods The SMMC-7721 cells were cultured with PIVKA-Ⅱ. The cell proliferation was assayed by cell counting kit-8 (CCK-8). The SMMC-7721 cell clone formation was assayed plate clone formation. The cell apoptosis was assayed by flow cytometry. The cell migration was assayed by transwell. The expression of transforming growth factor-α (TGF-α), basic fibroblast growth factor (bFGF), vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)-2 and MMP-9 were detected by Western blotting. Results The cell clone formation rate (1.94±0.14) and cell survival rate (1.53±0.21) in PIVKA-Ⅱ group was significantly higher than those in negative control group (1.12±0.08, 1.11±0.15) and blank control group (1.00±0.03, 1.00±0.04; F=68.254, 10.264, P=0.000, 0.000). The apoptosis rate of PIVKA-Ⅱ group(1.52±0.28) was lower than that of negative control group (3.13±0.36) and blank control group (3.45±0.43; F=48.436, P=0.000). The cell migration number (62.54±7.45) and mobility (2.13±0.21) of PIVKA-Ⅱ group was significantly higher than those in negative control group (31.34±6.03, 1.06±0.03) and blank control group (29.42±5.59, 1.00±0.05; F=46.241, 283.240, P=0.000, 0.000). The expression of TGF-α, bFGF, VEGF, MMP-2, MMP-9 (1.69±0.13, 1.30±0.04, 1.63±0.11) in PIVKA-Ⅱ group was higher than that in negative control group (1.11±0.07, 1.06±0.01, 1.08±0.02, 1.14±0.06, 1.17±0.04) and blank control group (1.00±0.01, 1.00±0.01, 1.00±0.02, 1.00±0.01, 1.00±0.01; F=2 014.254, 476.355, 1 845.364, 6.486, 7.045, P=0.000, 0.000, 0.000, 0.000, 0.000). Conclusion PIVKA-Ⅱ can promote the proliferation and migration of hepatocellular carcinoma cells and inhibit the apoptosis of hepatoma cells. The mechanism may be related to that PIVKA-Ⅱ can promote the expression of TGF-α, bFGF, VEGF, MMP-2 and MMP-9. Key words: Vitamin K deficiency or antagonist-Ⅱ-induced protein; Hepatocellular carcinoma; Proliferation