Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology
Article Title: Anti-cancer effects of fenbendazole on 5-fluorouracil-resistant colorectal cancer cells
Figure Lengend Snippet: Schematic representation of cell death pathways in colorectal cancer (CRC) cells following fenbendazole treatment. Fenbendazole induces G2/M arrest and apoptosis in both (A) 5-FU-sensitive SNU-C5 and (B) 5-FU-resistant SNU-C5 (SNU-C5/5-FUR) CRC cells. In SNU-C5 cells, fenbendazole is presumed to activate p53-mediated apoptosis by increasing p53 expression. In SNU-C5/5-FUR cells, fenbendazole triggers apoptosis without affecting p53 expression, whereas fenbendazole enhances ferroptosis by inhibiting the expression of GPX4 and SLC7A11. Therefore, although fenbendazole has anti-cancer effects on both 5-FU-sensitive and resistant CRC cells, the mechanism of action appears to be different. That is, fenbendazole promotes cell death by activating p53-mediated apoptosis in SNU-C5 cells, whereas by both enhancing p53-independent apoptosis and ferroptosis-augmented apoptosis in SNU-C5/5-FUR cells. PARP, poly (ADP-ribose) polymerase; HMGB1, high mobility group box 1; GPX4, glutathione peroxidase 4; LC3, light chain 3.
Article Snippet: Antibodies specific for c-Myc (9E10; #sc-40; diluted 1:1,000), cytochrome-C (H-104; #sc-7159; diluted 1:1,000), ERK (C-16; #sc-93; diluted 1:2,000), ferritin heavy chain (B-12) (FTH1; #sc-376594; diluted 1:2,000), GAPDH (#sc-47724; diluted 1:2,000), and phospho-p53 (Ser6; #sc135630; diluted 1:1,000), transferrin receptor (H68.4) (TfRC; #sc-65882; diluted 1:2,000) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA); glutathione peroxidase 4 (GPX4 [EPNCIR144]; #ab125066; diluted 1:1,000) and SLC40A1 (ferroportin, FPN; #ab78066; diluted 1:1,000) were purchased from Abcam (Cambridge, UK); caspase-3 (#9665; diluted 1:1,000), cleaved caspase-3 (#9661; diluted 1:1,000), cyclin B1 (#4138; diluted 1:1,000), phospho-ERK (Thr202/Tyr204; #4370; diluted 1:2,000), JNK (#9252; diluted 1:1,000), phosphor-JNK (Thr183/Tyr185) (#9251; diluted 1:1,000), p21 (#2947; diluted 1:1,000), p27 (#2552; diluted 1:1,000), p38 MAPK (#9212; diluted 1:1,000), phospho-p38 MAPK (Thr180/Tyr182; #9211; diluted 1:1,000), PARP (#9542; diluted 1:1,000), autophagy antibody sampler kit (#4445; consisting of Beclin-1 (D40C5), LC3 (D3U4C), Atg5 (D5F5U), Atg12 (D88H11), Atg16L1 (D6D5), Atg7 (D12B11), and Atg3; diluted 1:2,000/each), and necroptosis antibody sampler kit (#98110; consisting of receptor-interacting protein kinase (RIP) (D94C12), phospho-RIP (Ser166) (D1L3S), mixed lineage kinase domain-like protein (MLKL) (D2I6N), phospho-MLKL (Ser358) (D6H3V), receptor-interacting protein kinase 3 (RIP3) (E1Z1d), and phospho-RIP3 (S227) (D6W2T); diluted 1:1,000/each) were purchased from Cell Signaling Technology Inc. (Beverly, MA, USA); the high mobility group box 1 (HMGB1; #GTX62170; diluted 1:2,000) and p53 (#GTX70218; diluted 1:1,000) were purchased from GeneTex (Irvine, CA, USA); SLC7A11 (cysteine/glutamate transporter ; #ANT-111; diluted 1:2,000) was purchased from Alomone Labs (Jerusalem, Israel).