安五脂素对肺癌细胞的调控作用及机制OA

Objective To investigate the regulatory effect and underlying mechanism of anwulignan on lung cancer cells(A549).Methods CCK-8 assay was used to screen optimal administration concentration of anwulignan on A549 cells.Colony formation assay was performed to evaluate cell proliferation capacity.Transwell assay combined with scratch wound healing assay were employed for double validation of cell migration capacity.Annexin V-FITC/PI staining and Hoechst 33258 staining were performed to detect cell apoptosis.WST-1 assay was applied to determine levels of MDA,GSH and ROS.RT-qPCR was used to detect mRNA expression levels of BAX,CASP3,BCL2,PCBP2,SLC40 A1,TF and TFRC.Western blot was performed to analyze protein expression levels of apoptosis pathway,ferroptosis markers and Nrf2/HO-1 signaling pathway.Results When concentra-tion of anwulignan was higher than 10.0 μmol/L,it exerted a significant effect on proliferation of A549 cells(P<0.05,P<0.01),and could obviously inhibit cell proliferation and migration capacities(P<0.05,P<0.01).Compared with control group,apoptosis rate of cells in anwulignan treatment group was significantly increased.The expression levels of Caspase 3,Bax,SLC40A1 and TF proteins,as well as CASP3,BAX,SLC40 A1 and TF mRNAs,were all decreased(P<0.05,P<0.01).In contrast,the expression levels of Bcl-2,TFR1 and PCBP2 proteins,and BCL2,TFRC and PCBP2 mRNAs were all increased(P<0.05,P<0.01).Anwulignan could pro-mote production of MDA and ROS(P<0.05,P<0.01),reduce the content of GSH(P<0.05,P<0.01),and sig-nificantly up-regulate expression of Nrf2 and HO-1(P<0.01),as well as down-regulate expression of Keap1(P<0.05,P<0.01).Conclusion Anwulignan exerts an inhibitory effect on A549 cells,and its potential mechanism may be associated with the induction of ferroptosis and regulation of Nrf2/HO-1 signaling pathway,thereby affect-ing the processes of cell proliferation,apoptosis and migration.