Aspc 1

The human pancreatic cancer cell line AsPC-1, PaCa-2 and 4T1 breast cancer line were purchased from the Shanghai Cell Bank of the Chinese Academy of Sciences and cultured in Roswell Park Memorial Institute 1,640 (RPMI 1640) medium containing 10% (v/v) foetal bovine serum (FBS) and 1% penicillin/streptomycin. The cell lines were cultured using regular cell culture conditions (37°C with 5% CO₂).

Human pancreatic cancer cell lines consisting of AsPC-1, BxPC-3 CFPAC-1, L3.6pl, PANC-1, MIApaCa-2, SU.86.86, and one normal pancreatic cell line (HPNE) were purchased from the Shanghai Cell Bank, Chinese Academy of Sciences. The cells were cultured in Roswell Park MEMorial Institute (RPMI) 1640 combined with 10% fetal bovine serum (FBS; Gibco, Carlsbad, CA, USA). The L3.6pl cell line was cultured in Minimum Essential Medium (MEM) (Gibco, Grand Island, NY, USA) with 10% FBS (Gibco, Carlsbad, CA, USA). All cells were maintained at 37°C in 5% CO₂ incubator and passaged adopting standard cell culture techniques.

The immortalized human ovarian surface epithelial cells T29 and their transformed counterparts T29Kt1 (with the introduction of KRAS^V12 into the immortalized cells) were reported previously28 (link) and were cultured in medium consisting of 1:1 MCDB105 medium and M199 medium (Sigma-Aldrich Co.) with 10% fetal bovine serum (HyClone) in the presence of 1% penicillin and streptomycin (HyClone). HCT-116 (p53^+/+) and HCT-116 (p53^−/−) were kindly provided by Dr Chuangui Wang (East China Normal University, Shanghai, China) and were maintained in DMEM with the same supplements. The culture medium of other cells used is listed in Supplementary Table 2. A549, H441, H358, H1299, H1975, PC9, H661, H522, HCT-15, LoVo, SW480, SW620, DLD-1, HT29, CaCo2, HCT-8, CCD-18Co, CCD841CoN, Panc-1, CFPAC-1 and BxPC-3 were obtained from the American Type Culture Collection (Manassas, VA, USA); Calu-1, Calu-3, WI-38, MRC-5, T84, LS174T, SW1116 and AsPC-1 were obtained from the Shanghai Cell Bank of the Chinese Academy of Sciences (Shanghai, China). KRAS status identification by sequencing and limited genotyping in cancer cells was shown in Supplementary Table 2. Most of these cells were grown at 37 °C under a humidified 95:5 (%; v/v) mixture of air and CO₂. SW480 and SW620 cells were grown without CO₂. All of the cell lines were authenticated by short tandem repeat analysis.

Human normal pancreatic duct epithelial cell HPDE6-C7, and pancreatic cancer cell lines PANC-1, SW1990, AsPC-1 and BxPC-3 were purchased from Shanghai Cell Bank (Shanghai, China). DMEM medium (11,965,084, Gibco, Waltham, MA, USA) containing 10% fetal bovine serum (FBS, 10,099,141, Gibco) and 1% penicillin-streptomycin (15,070,063, Gibco) was used to culture cells in a humidified atmosphere at 37°C with 5% CO₂ [14 (link)].

Five human pancreatic ductal adenocarcinoma (PDAC) cell lines (PANC-1, BxPC-3, AsPC-1, CFPAC-1, and MIA-PaCa-2) and a normal human pancreatic ductal cell line (hTERT-HPNE) were purchased from the Shanghai Cell Bank (Shanghai, China). According to the recommendation of ATCC, both BxPC-3 and AsPC-1 cell lines were cultured in RPMI­1640 Medium (Life Technologies; Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS) (Life Technologies; Carlsbad, CA, USA) and 1% penicillin-streptomycin (HyClone, USA). However, PANC-1, CFPAC-1, and MIA-PaCa-2 were grown in Dulbecco's modified Eagle's medium (DMEM) (Life Technologies; Carlsbad, CA, USA) containing 10% FBS and 1% penicillin-streptomycin (HyClone, USA). In addition, the hTERT-HPNE cell line was maintained in DMEM, supplemented with 5% FBS, 10 ng/ml human recombinant epidermal growth factor (EGF), 5.5 mMD glucose (1 g/L), and 750 ng/ml puromycin. All cell lines were routinely cultured in a humidified incubator with 5% CO₂ at 37°C.

HEK293T, HPDE pancreatic duct epithelial cell line and human PAAD cell lines (AsPC-1, BXPC3, CFPAC1, PANC-1, and SW1990) cells were obtained from the Shanghai Cell Bank. Cells were cultured in DMEM media supplemented with 10% fetal bovine serum (FBS), 100 units/mL penicillin and 100 μg/mL streptomycin.