Activated Stellate Cell Paracrine HGF Exacerbated Pancreatic Cancer Cell Ferroptosis Resistance

Figure 1

The activated PSCs promote pancreatic cancer cell ferroptosis resistance. (a) A proposed model illustrating the coculture system of Panc02 cells. (b) Panc02 cells were treated with different concentrations of Erastin (0, 0.5, and 1 μM) or RSL3 (0, 0.05, and 0.1 μM) for 72 hours under normal or coculture and Fer-1 conditions. Cell viability was measured by CCK-8 kits. (c) qRT-PCR analysis of the mRNA expression levels of ferroptosis indicators (NRF2, SLC7A11, GPX4, FTH1, and NCOA4) in Panc02 cells under normal and coculture conditions. ACTB mRNA expression was detected as a loading control. (d) Western blot analysis of the protein expression levels of ferroptosis indicators (SLC7A11 and GPX4) in Panc02 treated with DMSO, coculture, Erastin (2 μM), and coculture+Erastin (2 μM). β-tubulin expression was detected as a loading control. (e–h) Panc02 cells were treated with DMSO, coculture, Erastin (2 μM)/RSL3 (0.1 μM), and coculture+Erastin (2 μM)/RSL3 (0.1 μM). Iron (Fe²⁺) level of Panc02 was evaluated by flow cytometry (e). The relative GSH (f) and MDA (g) concentrations of Panc02 were analyzed. Lipid ROS level of Panc02 was evaluated by flow cytometry (h). Fer-1 represents ferrostatin-1. Co-Panc02 represents Panc02 cells which were cocultured with activated PSCs. Experiments were repeated three times, and data were expressed as the . , , and .

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