Astragalus polysaccharide promotes ferroptosis and inhibits proliferation of hepatocellular carcinoma cells by regulating Wnt/β-catenin signaling pathway
ZHU Li-qing,WANG Zhi-xin,CAO Hu
Abstract:
Objective To investigate the regulatory effect of astragalus polysaccharide (APS) on ferroptosis mechanism of hepatocellular carcinoma. Methods HepG2 cells were treated with different concentrations of APS (0, 50, 100, 200 mg/L) for 48 hours, and were recorded as control group and APS low, medium and high dose group, respectively. Cell proliferation was detected with CCK-8 kit, while glutathione (GSH), Reactive Oxygen Species (ROS) and lipid peroxide levels of each group were detected with commercial kits, respectively. Flow cytometry was used to detect the iron level in each group. Glutathione Peroxidase 4 (GPX4) and long-chain acyl-CoA synthetase 4 (ACSL4), markers of ferroptosis, were detected by Western blot, as well as the expression levels of key proteins of the Wnt/β-catenin signaling pathway including Wnt and β-catenin. Results Compared with the control group, APS high-dose, medium-dose and low-dose groups could inhibit the proliferation of HepG2 cells, and the inhibitory effect was more significant with the increase of APS concentration (P<0.05). In addition, compared with the control group, the high, medium and low dose groups could reduce the GSH level in HepG2 cells, increase the ROS and lipid peroxide levels, and increase the iron level, and the effect was more significant with the increase of APS concentration (P<0.05). At the same time, Western blot further verified that compared with the control group, APS high, medium and low dose groups could reduce GPX4 protein expression level and increase ACSL4 protein expression level in a dose-dependent manner (P<0.05). Compared with the control group, the expression levels of Wnt and β-catenin protein in high, medium and low dose APS groups were decreased, and the inhibitory effect of protein expression was more significant with the increase of APS dose (P<0.05). Conclusion APS can enhance the accumulation of ROS and lipid peroxides, enhance ferroptosis and inhibit the proliferation of hepatocellular carcinoma cells, which may be related to the deactivation of Wnt/β-catenin signaling pathway.
