[Mechanism of artesunate-induced ferroptosis in triple-negative breast cancer cells through Wnt/β-catenin signaling pathway].

This article investigated the effect and mechanism of artesunate(ART) on ferroptosis in triple-negative breast cancer. Triple-negative breast cancer MDA-MB-231 cells and MDA-MB-468 cells were cultured in vitro. The effect of ART at different concentrations on the viability of MDA-MB-231 cells and MDA-MB-468 cells was detected by cell counting kit-8(CCK-8), and the appropriate treatment concentration of ART was screened out. The cells were randomly separated into a control group and groups of ART, ART + ferroptosis inhibitor Ferrostatin-1(Fer-1), ART + ferroptosis inhibitor deferiprone(DFP), and ART + Wnt/β-catenin agonist(SKL2001). The CCK-8 method was used to detect cell viability. Commercial kits were used to detect the release of lactate dehydrogenase(LDH) and the level of glutathione(GSH), superoxide dismutase(SOD), and malondialdehyde(MDA). Immunofluorescence was used to measure the level of reactive oxygen species(ROS) and intracellular Fe~(2+) and the protein expression level of glutathione peroxidase 4(GPX4) and β-catenin. Western blot was used to detect the protein expression of transferrin receptor 1(TFR1), divalent metal transporter 1(DMT1), light polypeptide(FTL), acyl-CoA synthetase long-chain family member 4(ACSL4), solute carrier family 7 member 11(SLC7A11), GPX4, β-catenin, and phosphorylated glycogen synthase kinase 3β(p-GSK-3β). The results showed that the survival rate of MDA-MB-231 cells and MDA-MB-468 cells decreased with increasing ART concentration. Compared with those of the control group, the LDH release rate and the levels of ROS, intracellular Fe~(2+), and MDA were significantly elevated(P<0.01), and the TFR1, DMT1, and ACSL4 protein expressions were significantly increased(P<0.01) in the ART group. The GSH and SOD levels were significantly reduced(P<0.01), and the FTL, SLC7A11, GPX4, β-catenin, and p-GSK-3β protein expressions were significantly decreased(P<0.05 or P<0.01) in the ART group. Compared with those of the ART group, the LDH release rate and the levels of ROS, intracellular Fe~(2+), and MDA were significantly decreased(P<0.05 or P<0.01), and the TFR1, DMT1, and ACSL4 protein expressions were significantly reduced(P<0.01) in the ART + Fer-1 group and the ART + DFP group. The GSH and SOD levels were significantly increased(P<0.05 or P<0.01), and the FTL, SLC7A11, and GPX4 protein expressions were significantly elevated(P<0.05 or P<0.01) in the ART + Fer-1 group and the ART + DFP group. After using SKL2001, the inhibitory effect of ART on the Wnt/β-catenin pathway was weakened. To sum up, ART inhibits the proliferation of triple-negative breast cancer cells by inducing ferroptosis, and its mechanism may be related to the regulation of the Wnt/β-catenin signaling pathway.