Aspc 1

The human PC cell line AsPC-1 was purchased from Cell Lines Service (CLS, Eppelheim, Germany) and cultured as described^{7 (link)}. Nitroxoline was purchased from Sigma (St. Louis, MO, USA). Cells were seeded in 100 mm culture dishes (2 × 10⁶ cells/dish) and the following day, cells were treated with 27 µM Nitroxoline, corresponding to the IC50 value for this drug in AsPC-1 cells^{7 (link)}, or with vehicle (Dimethyl Sulfoxide, DMSO) for 24 or 48 hours. Then, cells were dissociated with trypsin-EDTA 1X in Phosphate-Buffered Saline (PBS) and pelleted.

Human pancreatic cancer (PC) cell lines AsPC-1 and Capan-2 were purchased from Cell Lines Service (CLS, Eppelheim, Germany). PC cell line BxPC-3 was purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). The three cell lines are known to express the EGFR protein [20 (link), 21 (link)]. AsPC-1 and Capan-2 carry KRAS mutations, while BxPC-3 and AsPC-1 are TP53 mutated. Cells were cultured in RPMI 1640, supplemented with 10% FBS at 37 °C, 5% CO₂. Nelfinavir mesylate and erlotinib HCl (OSI-744) were obtained from Selleckchem (Munich, Germany). 8-hydroxy-5-nitroquinoline (nitroxoline) was purchased from Sigma (St. Louis, MO, USA).

Pancreatic cancer (PC) cell lines AsPC-1 and Capan-2 were acquired from Cell Lines Service (CLS, Eppelheim, Germany). AsPC-1 carries mutations both in KRAS and p53, while Capan-2 is KRAS mutated and p53 wild type. Cells were cultured in RPMI 1640 medium, supplemented with 10% fetal bovine serum (FBS) at 37 °C, 5% CO₂. Parbendazole, fenbendazole, mebendazole and oxibendazole were purchased from Sigma-Aldrich (St. Louis, MO, USA). Gemcitabine was obtained by Selleckchem (Houston, TX, USA). DMSO was used to prepare stock solutions of the different drugs. A fixed amount of DMSO was present in each experiment and the maximum percentage of the solvent in culture media (0.15%) was employed in drug combination experiments.