Levistolide A inhibits bladder cancer progression by suppressing GPX4 expression and activating ferroptosis

Bladder cancer is the second most common malignant tumor of the urinary system worldwide, with high incidence and mortality rates. However, existing drugs for treating bladder cancer often cause many adverse reactions. Levistolide A (LA), a natural compound isolated from the traditional Chinese herb Ligusticum chuanxiong Hort , has been identified as an anti-cancer agent. However, its role in bladder cancer and underlying mechanisms remain largely unknown. In this study, we used the MTT assay and Annexin V/PI staining to detect the co-induced death of bladder cancer cell lines 5637 and T24 by LA and different cell death inhibitors. The rescue experiment show that ferrostatin-1 (Fer-1, ferroptosis inhibitor) could rescue the proliferation of bladder cancer cells prevented by LA or not other inhibitors of cell death. At the same time, through in vivo verification using a xenograft model, LA can effectively inhibit the proliferation of bladder cancer cell line 5637. Subsequent RNA-seq analysis, nude mouse tumor model, MDA detection, and Western blotting, demonstrated that LA-enhanced lipid peroxidation by up-regulating PTGS2 and down-regulating GPX4, accompanied by dysregulation of intracellular iron homeostasis. Furthermore, we also found that in in vivo experiments, co-treatment with LA and RSL3 (ferroptosis inducer) increased the inhibitory effect on bladder cancer cells 5637, highlighting its potential for clinical application. This provides a novel therapeutic strategy for eliminating bladder cancer cells.