Generation of Gemcitabine-Resistant Pancreatic Cancer Cells

Human pancreatic cancer MIA PaCa-2 from the Bioresource Collection and Research Center (BCRC Number: 60139) in Taiwan was purchased. By sub-culturing MIA-PaCa-2 cells and gradually increasing gemcitabine doses from 50 to 500 nM for six months, gemcitabine-resistant MIA-PaCa-2 pancreatic cancer cells (MIA-GR100) were generated [26 (link),27 (link)]. MIR-GR100 and MIA PaCa-2 were cultured in MEM supplemented with 10% FBS, penicillin (100 U/mL), and streptomycin (100 μg/mL) at 37 °C with 5% CO₂. For inhibitor assays, MIA-GR100 cells were pre-treated with chloroquine (CQ, 100 mM), 3-Methyladenine (3-MA, 10 mM), and z-Val-Ala-Asp-fluoromethyl ketone (zVAD-FMK, 10 mM) at 37C for 1 h, and then HMJ-38 cells were treated for the indicated durations.