Bxpc 3

The human pancreatic cancer cell lines AsPC1, SW1990, BxPC‐3 and Panc‐1, and the normal human pancreatic ductal epithelial cell line HPDE6c7 were purchased from the National Collection of Authenticated Cell Cultures (Shanghai, China). The cells were incubated in a medium containing 10% foetal bovine serum and 100 U/mL penicillin and 100 μg/mL streptomycin (Sigma, St. Louis, MO, USA) in RPMI 1640 medium (Gibco, Rockville, MD) at 5% CO₂ at 37°C. For mild hypoxia treatment, cells were cultured under an atmosphere of c5% O₂–5% CO₂. The pcDNA‐HSPB1, pcDNA‐FUS and their negative control (Vector) were purchased from RiboBio Co., Ltd (Guangzhou, China) and transfected using RiboBio Transfection Kit (RiboBio Co., Ltd). Small interfering RNA against HSPB1 (si‐HSPB1), FUS (si‐FUS) and their negative control (scramble) were designed and synthesized by Invitrogen (Carlsbad, CA, USA). All transfections were performed by using Lipofectamine 3000 Transfection Reagent (Invitrogen) according to the manufacturer's instructions.

The human PC cell line MIApaca‐2, BxPC‐3, SW1990 and PANC‐1 cells were purchased from the National Collection of Authenticated Cell Cultures (Shanghai). PSCs were isolated from pancreatic tissues adjacent to PC tumours using the outgrowth method and immortalized as reported by Tuveson et al¹⁰ Cells were cultured as previously described.¹⁶ All patients signed informed consent.

Human pancreatic cancer cell lines PANC-1 and BxPC-3 were obtained from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). PANC-1 and BxPC-3 were cultured in Dulbecco's modified Eagle's medium (DMEM; HyClone Laboratories Inc., Logan, UT, USA) and Roswell Park Memorial Institute (RPMI)-1640 medium (HyClone Laboratories Inc., Logan, UT, USA), respectively, and supplemented with 10% (v/v) fetal bovine serum (FBS; Gibco, New York, NY, USA) and 1% (v/v) penicillin/streptomycin (P/S; Gibco, New York, NY, USA). Cultured cells were maintained at 37 °C with 5% CO2 in a humidified incubator.

Human PC cell lines (MIA PaCa-2, PANC-1, Capan-1, and BxPC-3) and two normal human pancreatic duct epithelial cell lines (HPDE6-C7 and HPNE) were obtained from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences in Shanghai, China. The cells were cultivated in DMEM (MD207-050, Gibco-BRL, USA) containing 10% fetal bovine serum (16000-044, Gibco, USA) and 1% penicillin–streptomycin solution (PR40022, Proteintech, Wuhan, China) at 37 °C with 5% CO₂. Cell lines underwent routine testing for mycoplasma every 3 months. The genetic identity of the cell lines was confirmed by short tandem repeat profiling.

The human PC cell lines PANC-1, BxPC-3 and ASPC-1 were purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). PANC-1 and ASPC-1 were maintained in DMEM media and BxPC-3 were cultivated in 1640 media (GIBCO, NY, USA). All media contained 10% heat-inactivated fetal bovine serum (FBS, GIBCO, USA), 100 U/mL penicillin and 100 μg/mL streptomycin (GIBCO, USA). All cell lines were incubated 37 °C in a 5% CO₂ atmosphere.