Journal: Translational Oncology
Article Title: Carfilzomib promotes Iodine-125 seed radiation-induced apoptosis, paraptosis, and ferroptosis in esophageal squamous cell carcinoma by aggravating endoplasmic reticulum stress
doi: 10.1016/j.tranon.2025.102393
Figure Lengend Snippet: CFZ enhances the pro-paraptotic effect of 125 I seed radiation in ESCC cells. (A) CFZ aggravated 125 I seed radiation-induced cytoplasmic vacuolation in Kyse-150 and EC-109 cells. Cells were treated with CFZ (20 nM, 24 h), 125 I seed radiation (6 Gy), and CHX (10 μM, 24 h) alone or in combination. Cell morphological changes were observed under light microscopy, and the percentage of vacuolated cells was measured. Scale bar (upper) = 100 μm, Scale bar (lower) = 30 μm. N = 6. (B and C) Combined treatment of 125 I seed radiation and CFZ increased cytosolic and mitochondrial Ca 2+ levels in Kyse-150 and EC-109 cells. Cells were treated with 125 I seed radiation (6 Gy) and CFZ (20 nM, 24 h) alone or in combination, then stained with Fluo-3 AM or Rhod-2 AM and quantitatively analyzed by mean fluorescence intensity using flow cytometry. (D) CFZ aggravated 125 I seed radiation-induced ubiquitination, ER stress, and UPR in Kyse-150 and EC-109 cells. Cells were treated with 125 I seed radiation (6 Gy) and CFZ (20 nM, 24 h) alone or in combination. Protein levels of Ub, β-actin (control for loading), Bip, p-eIf2α, IRE1α, ATF6, CHOP, and β-tubulin (control for loading) were analyzed by Western blot. (E) Combined treatment of 125 I seed radiation and CFZ did not influence the protein expression of Alix as analyzed by Western blot. N = 3, * P < 0.05, ** P < 0.01, and *** P < 0.001 compared to control group. # P < 0.05, ## P < 0.01, and ### P < 0.001 compared between indicated groups.
Article Snippet: Antibodies against p-H2AX (#9718), PARP (#9532), p53 (#2524), p-p53 (#9284), ubiquitin (Ub, #3936), p-eIF2α (#3398), IRE1α (#3294), ATF6 (#65,880), Alix (#92,880), LC3B (#3868), β-tubulin (#2128), β-actin (#4970) were purchased from Cell Signaling Technology (Danvers, MA, USA).
Techniques: Light Microscopy, Staining, Fluorescence, Flow Cytometry, Ubiquitin Proteomics, Control, Western Blot, Expressing
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