Gentiopicroside is an important plant secondary metabolite and has been reported to exhibit tremendous pharmacological potential. In the present study we investigated the anticancer effects of gentiopicroside against ovarian SKOV3 cancer cells.Anticancer effects were determined by MTT assay. Apoptosis was investigated by DAPI and annexin V/propidium iodide (PI) double staining. Mitochondrial membrane potential (MMP) determination and cell cycle analysis were carried out by flow cytometry. Protein expression was examined by western blotting.The results showed that gentiopicroside exerted anticancer effects on SKOV3 cancer cells in a dose-dependent manner. The IC50 of gentiopicroside was 20 µM against the SKOV3 cancer cells. The anticancer effects were mainly found to be due to loss of MMP and induction of apoptosis. The Bax/Bcl-2 expression ratio was also altered upon gentiopicroside treatment. Furthermore, gentiopicroside could arrest the SKOV3 cells in G2/M phase of the cell cycle and prevent their migration and invasion.These results indicate that gentiopicroside could prove a potential lead molecule in the treatment and management of ovarian cancer.