Figure 1.

Zalcitabine induces ALOX5-dependent ferroptosis in pancreatic cancer cells. (A) Upregulation of POLG gene expression within the tumor from PDAC patients compared to normal controls (*P < 0.05). (B) Western blot analysis of POLG protein expression in the indicated human PDAC or normal ductal cells. (C) The indicated human PDAC cells were treated with zalcitabine (2.5–20 µM) for 72 h, and the level of cell death was assayed (n = 3, *P < 0.05 versus the control group). (D) Heat map of levels of cell death in the indicated human PDAC cells following treatment with zalcitabine (20 µM) in the absence or presence of Z-VAD-FMK (20 µM), necrosulfonamide (1 µM), ferrostatin-1 (1 µM), or liproxstatin-1 (1 µM) for 72 h (n = 3). (E) Analysis of cell death in PANC1 cells following treatment with actinomycin D (1 µM) or HS-173 (1 µM) in the absence or presence of Z-VAD-FMK (20 µM) or necrosulfonamide (1 µM) for 72 h (n = 3, *P < 0.05). (F) Heat map of levels of gene expression in the indicated PDAC cells (n = 3). (G) Levels of gene expression in the indicated gene knockdown PDAC cells (n = 3, *P < 0.05 versus control shRNA group). (H, I) Analysis of levels of MDA (H) and cell death (I) in the indicated PDAC cells following treatment with zalcitabine (20 µM) for 72 h (n = 3, *P < 0.05 versus control shRNA group)