Norcantharidin induces ferroptosis via the suppression of NRF2/HO-1 signaling in ovarian cancer cells.

Abstract

Increasing evidence has indicated a crucial role of ferroptosis in ovarian cancer (OC). Norcantharidin (NCTD), a normethyl compound of cantharidin, is extensively used in clinical practice as an optional anticancer drug. However, whether NCTD leads to ferroptosis in OC has not been previously explored, at least to the best of our knowledge. In the present study, the effect of NCTD on SKOV3 and OVCAR-3 cells was evaluated. The experimental data of the present study revealed that NCTD significantly suppressed SKOV3 and OVCAR-3 cell viability in a concentration- and time-dependent manner. The results of Cell Counting Kit-8 assay revealed that NCTD treatment decreased SKOV3 and OVCAR-3 cell viability. In comparison, pre-incubation with ferrostatin-1 (Fer-1) significantly reversed the NCTD-induced reduction in SKOV3 and OVCAR-3 cell viability; however, no changes in cell viability were observed when the SKOV3 and OVCAR-3 cells were treated with NCTD, in combination with the apoptosis inhibitor, Z-VAD-FMK, the ferroptosis inhibitor, necrostatin-1, and the autophagy inhibitor, 3-methyladenine. Additionally, it was observed that NCTD markedly enhanced reactive oxygen species production and malondialdehyde and ferrous ion levels in the SKOV3 and OVCAR-3 cells; however, pre-incubation with Fer-1 abolished these effects. Flow cytometry also demonstrated a significant increase in cell death following treatment of the SKOV3 and OVCAR-3 cells with NCTD; however, pre-incubation with Fer-1 also reversed these effects. In vivo experiments demonstrated that NCTD significantly reduced tumor volume and weight. More importantly, it was revealed that nuclear factor erythroid 2-related factor 2 (NRF2), heme oxygenase 1 (HO-1), glutathione peroxidase 4 (GPX4) and solute carrier family 7 member 11 (xCT) expression levels were significantly decreased following NCTD treatment. Collectively, NCTD may represent a potent anticancer agent in OC cells, and NCTD-induced ferroptotic cell death may be achieved by inhibiting the NRF2/HO-1/GPX4/xCT axis.