Sw1116

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.

The human CRC cell lines HCT 116, LoVo, SW1116, and SW620 were acquired from Shanghai Cell bank of Chinese Academy of Sciences (Shanghai, China), apart from the human colon epithelial cell line FHC which were from Kunming Cell bank of Chinese Academy of Sciences (Kunming, China). The cells were cultured in the respective recommended media and culture conditions: McCOY's 5A (SIGMA, USA) + 10% fetal bovine serum (FBS, GIBCO, USA) and 5% CO₂ for HCT 116; Ham's F-12K Medium (Invitrogen, USA) + 10% FBS and 5% CO₂ for LoVo; Leibovitz's L-15 Medium (GIBCO, USA) + 10% FBS (GIBCO, USA) and 100% air for SW1116, and SW620; RPMI-1640 (GIBCO, USA) + 10% FBS (GIBCO, USA) for FHC. The culture temperature was 37 ℃.

The human colonic carcinoma cell lines LoVo, CaCo2, SW1116, SW480 and HCT-116, and the normal colonic mucosa cell line NCM460 were purchased from the Shanghai Cell Bank. All cell lines were cultured in DMEM medium containing 10% FBS, 100 U/ml penicillin and 100 mg/ml streptomycin at 37 °C in a 5% CO₂ incubator.