Plantago depressa Willd. is a distributed herb which traditionally used for treating wound, fever, cancer and inflammation in some temperate asian countries. This present study aimed to investigate the anti-cancer activity of ethanol extract of Plantago depressa Willd. plant on B16F10 skin cancer cells. Cells were treated with different concentration of the PDE (25, 50, 100, 200 μg/mL) with or without lipopolysaccharide (LPS) stimulation to evaluate its effect on cell viability. Furthermore, anti-cancer activity was evaluated by checking viability of B16F10 skin cancer cells in various concentration of the PDE extract (0.6, 1.2, 2.5 and 5 μg/mL), using CCK-8 assay. The results indicated that (i) the PDE treatment did not influence on cell viability of HaCaT skin normal cells dose-dependently; and (ii) PDE exhibits anti-cancer activity through inhibition of proliferation of B16F10 skin cancer cells.