Ncm460 cell line

The human-derived nontransformed colonic epithelial cells line NCM460 cell line [52 (link)] was obtained from INCELL (San Antonio, TX, USA) and maintained as a monolayer in RPMI 1640 medium (Gibco) supplemented with 10% newborn calf serum (Gibco), 1% penicillin (5000 IU/mL)/streptomycin (5 mg/mL) solution (Gibco), 1% l-glutamine (2 mM) (Sigma, St. Louis, MO, USA), and cultured at 37 °C and 5% CO₂ in a humidified incubator. NCM460 cells were subcultured at a density of 1 × 10⁵/mL into 24-well plates or 6-well plates and cultured in RPMI1640 medium containing 0, 20, 40, 80 or 160 µg/mL PE for 72 h.

The malignant melanoma cell line G-361, human colorectal cancer cell lines HT-29 and HCT116 (American Type Culture Collection, Manassas, VA, USA Cat. No. CRL-1687, CCL-218 & CCL-247, respectively) were cultured with McCoy's Medium 5a (Gibco BRL, VWR, Mississauga, ON, Canada) supplemented with 10% (v/v) FBS (Thermo Scientific, Waltham, MA, USA) and 40 mg/ml gentamicin (Gibco, BRL, VWR). The ovarian adenocarcinoma cell line OVCAR-3 (American Type Culture Collection, Cat. No. HTB-161) was cultured in RPMI-1640 media (Sigma-Aldrich Canada, Mississauga, ON, Canada) supplemented with 0.01 mg/mL bovine insulin, 20% (v/v) fetal bovine serum (FBS) standard (Thermo Scientific, Waltham, MA, USA) and 10 mg/mL gentamicin. The pancreatic adenocarcinoma cell line BxPC-3 (American Type Culture Collection, Cat. No. CRL-1424) was cultured in RPMI-1640 medium, supplemented with 10% (v/v) fetal bovine serum (FBS) standard and 40 mg/mL gentamicin. Normal-derived colon mucosa NCM460 cell line (INCELL Corporation, LLC., San Antonio, TX, USA) was grown in INCELL's M3Base medium (INCELL Corporation, LLC., Cat. No. M300A500) supplemented with 10% (v/v) FBS and 10 mg/mL gentamicin.
All cells were grown in optimal growth conditions of 37°C and 5% CO2. Furthermore, all cells were passaged for ≤6 months.

We have used the normal-like human colon epithelial cell line (NCM460D) and three isogenic human CRC cell lines: HCT116 wild-type for p53 (HCT116), p53 null (HCT116 p53^−/−), and 5-FU-resistant HCT116 (HCT116-5FUR). The NCM460 cell line was purchased from INCELL Corporation, LLC San Antonio, TX, USA. The HCT116 cell line was obtained from the American Tissue Culture Collection, ATCC, Manassas, VA, USA. The HCT116 p53^−/− cells were provided by Carlos Maria Galmarini, PharmaMar, Madrid, Spain. The HCT116-5FUR cell line was generated in our laboratory from the parental HCT116 cells [18 (link)]. NCM460D cells were cultured in M3:Base media (INCELL) supplemented with 10% fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA). HCT116 and HCT116-5FUR cells were cultured in an RPMI 1640 (Lonza, Basel, Switzerland) medium supplemented with 10% FBS (Sigma-Aldrich), 100 U/mL of penicillin-streptomycin antibiotics (Lonza), and 1 mM of sodium pyruvate solution (Lonza). HCT116 p53^−/− cells were cultured in a DMEM medium (Lonza) supplemented with 10% FBS, 100 U/mL of penicillin-streptomycin, 1 mM of sodium pyruvate, and 1× MEM non-essential amino acid (Sigma-Aldrich). All cells were incubated in a humidified incubator (95% air, 5% CO₂) at 37 °C.