Erlotinib hcl osi 744

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).

The human pancreatic cancer cell lines AsPC-1, BxPC3, Capan-1, Capan-2, CFPAC-1, HS766T, Panc-1, and SW1990 were purchased from American Type Culture Collection (Manassas, VA) and normal human pancreatic duct epithelial (HPDE) cells were isolated from normal pancreatic tissues as described [41 (link)]. HPDE, Capan-1, Capan-2, HS766T, and Panc-1 were maintained in Dulbecco's modified Eagle medium (DMEM) with 10% FBS (Gibco, Carlsbad, CA). AsPC-1, BxPC-3, HPAF, SW1990 and HUVEC cell lines were maintained in RPMI 1640 with 10% FBS. CFPAC-1 was maintained in IMDM (Iscove's modified Dulbecco's medium) with 10% FBS. Human pancreatic stellate cells (PSCs) were purchased from ScienCell research laboratory (Carlsbad, CA) and maintained in stellate cell medium (ScienCell). The authenticity of the cells was determined by short tandem repeat analysis technology (Cell ID^™ System, Promega, Madison, WI). All cells were cultured at 37°C in an atmosphere of 5% CO₂ in air. Human recombinant periostin protein (rPeriostin) was purchased from Biovendor (Heidelberg, Germany) and dissolved in 0.1 M acetate buffer (pH 4) at a concentration of 1 μg/mL. The EGFR inhibitor Erlotinib HCl (OSI-744) and the Erk inhibitor SCH772984 were purchased from Selleck Chemicals (CA, USA), dissolved in 100% DMSO at 10 mM, and stored at −20°C. The final DMSO concentration in the medium was < 0.1% for all experiments.