Hs 578bst

Pancreatic cancer cell lines AsPC-1, Capan-2, and Panc-1 were obtained from the American Type Culture Collection (Rockville, MD). The cells were cultured in RPMI 1640 medium supplemented with 10% fetal calf serum, penicillin (100 U/ml), and streptomycin (100 μg/ml) in a humidified incubator containing 5% CO₂ at 37°C. The normal human breast epithelial cell line Hs 578Bst and normal human liver cell line L02 were purchased from Shanghai Cell Bank (Shanghai, China). These cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal calf serum, penicillin (100 U/ml), and streptomycin (100 μg/ml) in a humidified incubator containing 5% CO₂ at 37°C. AsPC-1 and Capan-2 cells were serum starved for 4–6 h before stimulation with EGF at a final concentration of 50 ng/ml for 15 min. Tissue samples were collected from patients during pancreatic resections for PDAC (n = 46). Normal pancreatic tissue samples were obtained through an organ donor procurement program when there was no suitable recipient for pancreatic transplantation (n = 11). Pancreatic tissues were immediately stored at –80°C or formalin-fixed and paraffin-embedded for histological analysis. The use of human tissue was approved by the local ethics committee (Tongji Medical College, China) and written informed consent was obtained from patients prior to surgery.

A normal human breast epithelial cell line Hs 578Bst and four breast cancer cell lines (MDA-MB-415, MDA-MB-231, MDA-MB-468 and MCF7) were purchased from Shanghai Cell Bank, Chinese Academy of Sciences (Shanghai, China). All the cells were cultured in DMEM (Gibco, Grand Island, New York, U.S.A.) supplemented with 10% fetal bovine serum (FBS, Sigma–Aldrich, St. Louis, MO, U.S.A.). Cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂.