Ccd 18co

Human colorectal cancer cell line HCT116, HT-29 and Colo205 were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA). SW480, SW620, Caco-2 and CCD-18co were kindly gift from Min-Chuan Huang (Graduate institute of Anatomy and Cell Biology, College of Medicine, National Taiwan University) and Kuen-Haur Lee (Program for Cancer Biology and Drug Discovery, College of Medical Science and Technology, Taipei Medical University). HCT116, HT-29 and Colo205 were cultured in RPMI-1640 medium (Gibco, Invitrogen, Carlsbad, CA, USA); SW480, SW620 and Caco-2 were maintained in DMEM medium (Gibco, Invitrogen, Carlsbad, CA, USA); CCD-18co was in MEM medium (Gibco, Invitrogen, Carlsbad, CA, USA). All of them were supplemented with 10% heat-inactivated FBS, penicillin 100 U/ml, streptomycin 100 μg/ml, and amphotericin B 2.5 μg/ml. Cells were maintained at 37°C in a humidified incubator with 5% CO₂/95% air. Hypoxia incubations were performed in hypoxia chamber, which was flushed with 1% O₂, 5% CO₂, and 94% N₂ at 37°C (BioSpherix Proox 110), and incubated for the indicated time.

A human colon fibroblasts cell line CCD-18Co and four CRC cell lines including HT29, SW480, HCT116, and Caco-2 were purchased from American Type Culture Collection (ATCC, Rockville, MD). CCD-18Co cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) with high glucose supplemented with 10% to 20% fetal bovine serum (FBS, Invitrogen, Carlsbad, CA, USA), 1% sodium pyruvate (100 mM), 1% non-essential amino acids (10 mM), and 1% penicillin/streptomycin. 1% sodium pyruvate (100 mM). HT29 and HCT116 cells used McCoy’s-5a–modified medium (Gibco, Grand Island, NY, USA), SW480 used RPMI 1640 medium (Invitrogen, Carlsbad, CA) and Caco-2 used Leibovitz’s L-15 Medium (Gibco), supplemented with 10% to 20% of FBS and 1% penicillin/streptomycin. Cells were maintained at 37°C with 5% CO₂ and humidified air.

The RD (human embryonal rhabdomyosarcoma) and CCD-18Co (human colon fibroblast) cell lines were purchased from the Korean Cell Line Bank (KCLB). SJCRH30 (human alveolar rhabdomyosarcoma) cell line was purchased from the American Type Culture Collection (ATCC). RD and CCD-18Co cells were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin/streptomycin (P/S; Gibco). SJCRH30 cells were maintained in Roswell Park Memorial Institute medium (RPMI 1640, Gibco) supplemented with 10% FBS and 1% P/S. Cells were grown in a humidified chamber containing 5% CO₂ at 37°C. Cell line authentication was performed by short tandem repeat profile using PowerPlex® 18D system (Promega Corporation) and sequencing apparatus Applied Biosystems 3130xl Genetic Analyzer (Applied Biosystem) according to manufacturer’s protocol.