基于Nrf2/SLC7A11/GPX4通路探讨清解扶正颗粒诱导结直肠癌细胞铁死亡的实验研究OA

Objective To investigate the in vitro mechanism of colorectal cancer(CRC)HCT-116 ferroptosis in cells induced by Qingjie Fuzheng Granules(QFG)through the signaling pathways involving Nuclear Factor E2-related Factor 2(Nrf2)/Solute Carrier Family 7 Member 11(SLC7A11)/Glutathione Peroxidase 4(GPX4).Methods Human colorectal cancer cell line HCT-116 was cultured in vitro and treated with different concentrations of QFG(0,0.05,0.10,0.25,0.50,0.75,1.00,1.50,and 2.00 mg/L).The inhibitory effect of QFG on cell proliferation was detected by CCK-8 method.According to the results,the doses of 0,0.5,1,and 2 mg/mL were designated as the control,low-,medium-,and high-dose groups respectively.Ultrastructural observations were performed via transmission electron microscopy;quantitative analysis of iron ion levels,reactive oxygen species(ROS),superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione(GSH)were measured using specific kits.Western blot was employed to detect protein expression levels of Nrf2,SLC7A11,and GPX4.Results QFG exhibited dose-dependent inhibition of HCT-116 cell proliferation.Transmission electron microscopy revealed that after QFG treatment,cells exhibited typical morphological features of ferroptosis,such as shrunken mitochondria and ruptured cristae.QFG dose-dependently increased the levels of iron ions,ROS,and MDA(P<0.05,P<0.01),while decreased the activity of SOD and GSH(P<0.01).Meanwhile,QFG downregulated Nrf2,SLC7A11,and GPX4 protein expression(P<0.01),and this inhibitory effect was more pronounced with increasing concentrations.Conclusion QFG may induce ferroptosis in HCT-116 cells by inhibiting the Nrf2/SLC7A11/GPX4 signaling pathway,thereby exerting an anti-CRC effect.