Phytochemicals play a prominent role as cancer preventive agents. Stevioside is a phytochemical which is isolated from Stevia rebaudiana and is known for its medicinal properties including prevention against cancer. Therefore, the present work intended to explore the antiproliferative activity of stevioside on ovarian cancer cell line, OVCAR-3. The results elucidated that stevioside treatment repressed the growth of OVCAR-3 cells and induced cytotoxicity, both dose dependently and time dependently. Furthermore, stevioside has been found to be associated with increased ROS production in the cell which suggests the initiation of apoptosis. Additionally, the decrease in mitochondrial membrane potential indicated the involvement of stevioside induced intrinsic apoptotic pathway in the cell. These results were further confirmed by increased level of caspase-3 and caspase-9 in OVCAR-3 cancer cells after stevioside treatment. Moreover, flow cytometric study established the apoptotic behavior of stevioside and showed cell cycle arrest in G2/M phase. Additionally, the inactivation of P13K/AKT signaling pathway was also revealed. Thus, the results may summarize the therapeutic potential of stevioside against ovarian cancer and its role as chemotherapeutic agent in future clinical applications.