Ls180

The human colon adenocarcinoma line LS174T was kindly provided by Dr. HA Choudry (University of Pittsburgh Medical Center, Pittsburgh, PA, USA) and LS180 was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The human colorectal carcinoma HCT116 wild type was obtained from Dr. B Vogelstein (Johns Hopkins University) and CX-1 cell line was obtained from Dr. JM Jessup (National Cancer Institute). Cell lines were maintained in Dulbecco’s Modified Eagle Medium (DMEM, Gibco, Gaithersburg, MD, USA), McCoy’s 5A or RPMI 1640 supplemented with 10 % fetal bovine serum (FBS, Atlanta Biological, Flowery Branch, GA, USA), 100 U/mL penicillin and 100 µg/mL streptomycin (Gibco) in a 5 % CO₂ incubator. These adherent cells, except for LS180, were subcultured every 3–4 days by treatment with a trypsin–EDTA solution (Gibco). The LS180 cell line was cultured by following a procedure of ATCC subculturing.

Established human colon cancer cell lines (LS180, HCT116, and HT29) were purchased from European Collection of Cell Cultures (ECACC) and immediately stored in liquid nitrogen. Cells used for specific investigations were recovered from cryopreserved aliquots and cultured for a maximum of 10 passages. LS180 and HCT116 were maintained in RPMI-1640 supplemented with 10% FBS, GlutaMAX-I, nonessential amino acids (NEAA), 100 mM sodium pyruvate, 10 mM HEPES (all from GIBCO), and 50 mM 2-mercaptoethanol (Sigma-Aldrich, St. Louis, MO, USA), whereas HT29 was maintained in McCoy’s 5A medium (Sigma) supplemented with 10% FBS and GlutaMAX-I.
Absence of mycoplasma contamination in cultured cells was verified by PCR testing prior to investigation. In specific experiments, LS180 cells co-expressing green fluorescent protein (GFP) and firefly luciferase (GFP/Luc-LS180 cells) [27 (link)] were also used.