Fig. 5. Fenbendazole-induced ferroptosis via glutathione peroxidase 4 (GPX4) in colorectal cancer cells.
(A) Effect of fenbendazole on the expression of ferroptosis-related proteins in SNU-C5 and SNU-C5/5-FUR cells was detected by immunoblotting. SNU-C5 and SNU-C5/5-FUR cells were mock-treated with dimethyl sulfoxide (DMSO) or treated with indicated dose of fenbendazole for 3 days. Total protein was collected, and immunoblotting analysis was performed for GPX4, high mobility group box 1 (HMGB1), SCL7A11, ferritin heavy chain (FTH1), ferroportin (FPN), and transferrin receptor (TfRC). GAPDH was used for a loading control. Band density was analyzed by AzureSpot analysis software, and results are expressed as the mean ± SD. *p < 0.05, **p < 0.01 vs. DMSO. (B) The SNU-C5 and SNU-C5/5-FUR cells were treated with fenbendazole alone or with fenbendazole in combination with indicated doses of ferrostatin-1 or deferoxamine mesylate (DFOM) for 3 days. The extent of cell viability was determined by MTT assay. Data are presented as mean ± SD.
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