Glycycoumarin promotes ferroptosis in human breast cancer cells through NRF2/HO-1 signaling axis
HE Miao,XU Jing,LIU Bin
Abstract:
Objective To investigate how glycycoumarin (GCM) promotes ferroptosis in human breast cancer cells via the nuclear factor erythroid 2-related factor 2 (NRF2)/heme oxygenase-1 (HO-1) signaling pathway. Methods BC cell lines (MCF-7 and MDA-MB-231) were cultured with different concentrations (25, 50, 75 μmol/L) of GCM for 24 hours, and the inhibitory effect of GCM on the proliferation of BC cells was evaluated by CCK-8 method and colony formation assay. Transwell and wound healing assay were used to detect the inhibitory effect of GCM on invasion and migration of BC cells. Flow cytometry was used to detect the promoting effect of GCM on apoptosis of BC cells. ELISA was used to detect the effect of GCM on ferroptosis-related indexes of BC cells, and Western blot was used to detect the expression of ferroptosis-related proteins. Results Compared with the control group (DMSO), the experimental groups (25, 50, 75 μmol/L GCM) significantly reduced the proliferation of BC cells to 80%, 73%, and 63% at 25, 50, and 75 μmol/L, respectively, and significantly inhibited colony formation (all P <0.05). It also significantly inhibited the invasion and migration of BC cells (both P <0.05). GCM significantly promote the apoptosis of BC cells, and the apoptotic proportion increased with increasing GCM concentration (all P <0.01). After treatment with 75 μmol/L GCM, the most significant effect on the viability of BC cells was observed with Ferrostatin-1 (P <0.001). GCM promoted ferroptosis-related indices and related protein levels in BC cells, and increased Fe2+, malondialdehyde, reactive oxygen species levels and acyl-CoA synthetase long-chain family member 4 protein levels, and decreased glutathione level, solute carrier family 7 member 11 and glutathione peroxidase 4 protein levels (all P <0.05). In addition, GCM promoted the expression levels of NRF2 and HO-1 in BC cells (P <0.05). These results suggest that GCM promotes ferroptosis in BC cells by activating NRF2/HO-1 signaling axis. Conclusion GCM promotes ferroptosis of human breast cancer cells by activating NRF2/HO-1 signaling axis, thereby inhibiting cell viability, proliferation, invasion and migration.
