Effects of Methionine Restriction on the Proliferation,Apoptosis,and Ferroptosis of Osteosarcoma Cells
Objective:To investigate the effects of methionine restriction on the proliferation,apoptosis,and ferroptosis of os-teosarcoma cells.Methods:Three human osteosarcoma cell lines,MNNG-HOS,U2OS and Saos-2,were treated with normal or methion-ine-restricted medium,and the effects of methionine restriction on cell growth were assessed by cell counting;High-throughput sequenc-ing(RNA-seq)was employed to analyze the molecular changes in osteosarcoma cells following methionine restriction;Flow cytometry was used to evaluate the cell cycle,cell apoptosis,reactive oxygen species(ROS),and lipid ROS levels;Western blot analysis was con-ducted to determine the protein expression of key ferroptosis biomarkers GPX4 and COX2.Results:Methionine restriction significantly inhibited the proliferation of osteosarcoma cells(P＜0.001);RNA-seq analysis identified 1719 differentially expressed genes.The enrich-ment analysis indicated that the Ferroptosis pathway was significantly activate,and cell cycle pathway were significantly inhibited(P＜0.001);Cytological experiments showed that methionine restriction induced G2/M Cell Cycle arrest and apoptosis in osteosarcoma cells;Besides,methionine restriction could induce ROS,lipid ROS level,and decrease GPX4 protein expression,upregulate COX2 protein ex-pression in osteosarcoma cells.Moreover,methionine restriction Induced ferroptosis could be partially rescued by ferrostatin-1.Conclu-sions:Our findings provide potential novel therapeutic approaches or combination therapies for treating osteosarcoma while offering a theoretical basis for utilizing methionine restriction therapy against osteosarcoma.
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