Study Of The Role Of DL-Buthionine-(S,R)-Sulfoximine In Causing Iron Death In Colon Cancer Cells

Objective: The effect of DL-Buthionine-(S,R)-sulfoximine(DL-BSO)in ferroptosis in colon cancer cells was investigated.Methods: DL-BSO at concentrations of 100,200,300,400 and 500μmol-L-1 was used to interfere with colon cancer HCT-116 cells and SW480 cells,and the cells were cultured in 96-well plates for 24,36 and 48 H.The change of cell viability was detected by Cell Counting Kit-8(CCK-8),and DL-BSO at 300 μmol-L-1 was used to interfere with colon cancer HCT-116 cells and SW480 cells.The cells were randomly divided into control group and DL-BSO group,and the transwell assay and scratch assay were used to detect the lateral and longitudinal migration ability of colon cancer cells.and necrosis inhibitor Nec-1 and iron death inhibitor Fer-1 both at 5μmol/L pretreatment,were divided into control group,DL-BSO group,inhibitor group,DL-BSO combined with inhibitor group,and cell viability changes were detected by Cell Counting Kit-8(CCK-8),and DL-BSO at a concentration of 300μmol-L-1 was used to intervene colon cancer SW480 and HCT-116 cells,divided into control and DL-BSO groups,and the expression levels of glutathione peroxidase 4(GPX4)and cystine-glutamate reverse transporter subunit x CT protein were detected by protein blotting.300μmol-L-1concentration of DL-BSO was used to intervene in colon cancer SW480 and HCT-116 cells,divided into control and DL-BSO groups The level of ROS was detected by fluorescence development of lipid peroxidation sensor C11-BODIPY-591,and colon cancer cells were pretreated with 5μmol/L Fer-1,followed by intervention with DL-BSO at a concentration of 300μmol-L-1,divided into control,DL-BSO,inhibitor,and DL-BSO combined with inhibitor groups,and iron was detected by iron ion colorimetric kit ion concentration,glutathione concentration by glutathione kit,(GPX4),x CT expression level by protein blotting,and ROS level by lipid peroxidation sensor C11-BODIPY-591 fluorescence development and loss cytometry.Results: After DL-BSO intervention,cell activity was significantly reduced,and the optimal concentration and treatment time of DL-BSO were 300μmol L-1 and 36H;cell lateral and longitudinal migration ability were affected;GPX4 and XCT,probe detection and loss cytometer showed increased ROS level,iron ion concentration,glutathione concentration,and DL-BSO was not affected by Nec-1 but affected by Fer-1.Conclusion: Butycythiosulfonyimide(DL-BSO)can cause iron death in colon cancer cells by inducing the production of ROS,which can be used as one of the anti-tumor effects of butathiosulfonyimide.