Ovarian cancer (OC) is a common malignant tumor with the greatest mortality rate among gynecological tumors. Tiliroside (TIL) is a glycosidic dietary flavonoid with various pharmacological activities. The purpose of this study was to investigate the exact mechanism by which TIL eliminates OC cells. In vitro, TIL exerted anti-tumor activities by inducing cell death and inhibiting the invasion and migration of A2780 and OVCAR8 cells. Additionally, the suppressive effect of TIL on OC cells was mainly due to the induction of ferroptosis, as demonstrated by the fact that only ferroprostatin-1 (Fer-1) significantly inhibited the anti-tumor activity of TIL, with the accumulation of ROS, MDA, and Fe Our research revealed that TIL simultaneously targets AKR1B1 and modulates iron metabolism, thereby inducing ferroptosis and improving anti-tumor efficacy. As a novel drug, TIL is promising for OC treatment.