Theaflavin-3,3-Digallate Plays a ROS-Mediated Dual Role in Ferroptosis and Apoptosis via the MAPK Pathway in Human Osteosarcoma Cell Lines and Xenografts

Figure 3

TF3 induced oxidative stress and altered iron metabolism in OS cell lines. OS cells were treated with 0.05% DMSO (vehicle control) or the indicated concentrations (0, 40, 80, 120, or 160 μM) of TF3 for 24 h. (a) Representative fluorescence images of MG63 and HOS cells stained with DCFH-DA (green). (b) Statistical graphs of ROS levels detected with a fluorescence reader in MG63 and HOS cells stained with DCFH-DA (green). (c) Statistical graphs of GSH levels detected with a GSH assay kit in MG63 and HOS cells. (d) Statistical graphs of MDA levels detected with an MDA assay kit in MG63 and HOS cells. (e, g) Representative fluorescence images of cells stained with FerroOrange and Hoechst 33342 solution in the TF3 treatment and control groups. (f, h) Statistical graphs of Fe²⁺ levels detected by a fluorescence reader in MG-63 and HOS cells stained with FerroOrange solution. (i) Western blotting and statistical analyses of ferroptosis-related proteins, including FTH and GPX4, in MG63 and HOS cells. The data are presented as the (). , , , and vs. the vehicle control group.

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