小白菊内酯对人非小细胞肺癌A549细胞铁死亡的影响及其作用机制研究OA

Objective To investigate the effect of parthenolide(PN)on ferroptosis in human non-small cell lung cancer A549 cells and its mechanism of action.Methods A549 cells were treated with different concentrations of PN and deferoxamine(DFO),and CCK-8 assay was used to measure cell viability.A549 cells were divided into groups A,B,C,and D.The cells in group A were not given any drug treatment.The cells in group B were treated with PN(30 μmol/L).The cells in groups C and D were pretreated with DFO(50 μmol/L)for 1 hour;then,the cells in group C were further treated with DFO(O50 μmol/L),while those in group D were treated with PN(30 μmol/L)+DFO(50 μmol/L).Transmission electron microscopy was used to observe mitochondrial structure;JC-1 assay was used to measure mitochondrial membrane potential;DCFH-DA staining,ferrous iron fluo-rescence assay,thiobarbituric acid assay,and 5,5'-dithiobis(2-nitrobenzoic acid)assay were used to measure the levels of reactive oxygen species(ROS),Fe2+,malondialdehyde(MDA),and glutathione(GSH)in each group;Western blot was used to measure the protein relative expression levels of DRP1,SLC7A11,GPX4,TFR1,and FSP1 in each group.Results CCK-8 assay showed that A549 cell viability gradually decreased with the increase in PN concentration,and DFO could reverse this effect(F=144.80-4 287.00,q=11.68-57.47,P＜0.05).Transmission electron microscopy showed that group B exhibited marked mito-chondrial shrinkage and loss of cristae structure,accompanied by cell membrane rupture and vacuolization,and group D showed significant improvements in mitochondrial morphology compared with group B.Measurement of mitochondrial membrane potential showed that group B had a significantly lower mitochondrial membrane potential than group A,and groups C and D had a significantly higher mitochondrial membrane potential than group B(F=514.70,q=42.75-46.68,P＜0.05).Biochemical analyses showed that compared with group A,group B had a significantly lower level of GSH and significantly higher levels of Fe2+,ROS,and MDA,and compared with group B,group D had a significantly higher level of GSH and significantly lower levels of Fe2+,ROS,and MDA(F=57.24-11 646.00,q=14.88-218.70,P＜0.05).Western blot showed that compared with group A,group B had significantly higher protein expression levels of DRP1 and TFR1 and significantly lower protein expression levels of GPX4 and SLC7A11,and compared with group B,group D had significantly lower protein expression levels of DRP1 and TFR1 and a signifi-cantly higher protein expression level of SLC7A11(F=160.60-14 969.00,q=5.68-278.30,P＜0.05).Conclusion PN can in-duce ferroptosis in A549 cells,possibly by inhibiting signal transduction along the SLC7A11-GSH-GPX4 axis.