Sw480

Human colorectal cancer cell lines (HCT116, SW480 and LoVo), human breast cancer cell lines (MCF-7 and MDA-MB231) and human hepatocellular carcinoma (HepG-2) were purchased from Korean Cell Line Bank (Seoul, Korea) and grown in DMEM/F-12 supplemented with 10% fatal bovine serum (FBS), 100 U/ml penicillin and 100 μg/ml streptomycin. The cells were maintained at 37°C under a humidified atmosphere of 5% CO₂. The extracts of ginger leaf (GL) were dissolved in dimethyl sulfoxide (DMSO) and treated to cells. DMSO was used as a vehicle and the final DMSO concentration did not exceed 0.1% (v/v).

Human colorectal cell lines, including HCT116, HT29, SW480, and SW620, were purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea). Each was propagated in RPMI1640 (ThermoFisher, Waltham, MA, USA) and supplemented with 10% fetal bovine serum (FBS, Thermofisher) and 1% penicillin/streptomycin (Gibco by life technologies, Thermo Fisher Scientific, Waltham, MA, USA)) in a fully humidified incubator containing 5% CO₂ at 37 °C.

HeLa (cervical cancer, Korea Cell Line Bank, Seoul, Korea) and SW480 (colon cancer, Korea Cell Line Bank) cells were incubated in cell media containing DMEM 89%, FBS 10%, and 0.1 mg/mL of an antibiotic-antimycotic agent. The cells were incubated in a cell incubator (37 °C, 5% CO₂).

The human CRC cell lines Caco2, SW480, HT29, HCT116, LoVo, and SW48 were obtained from Korean Cell Line Bank (KCLB, Seoul, Korea) or American Type Culture Collection (ATCC, Manassas, VA, USA). SW480, SW48, HCT116, LoVo, and HT29 cells were cultured at 37 °C in Roswell Park Memorial Institute Medium (RPMI) 1640 (HyClone, Logan, UT, USA), including 10% fetal bovine serum (FBS) in a humidified atmosphere of 5% CO₂. Caco2 cells were cultured at 37 °C in a-MEM (HyClone), including 20% FBS in a humidified atmosphere of 5% CO₂.

Human colon cancer cell lines HT29, HCT116, SW480, and SNU-C5 were obtained from the Korean Cell line Bank (Seoul, Korea). HT29, HCT116, SW480, and SNUC5 cells were cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum (FBS). All cell cultures were incubated at 37 °C in a 5% CO₂ humidified incubator. The primary antibodies used included Poly-(adenosine diphosphate-ribose) polymerase (Cell Signaling Technology, Danvers, MA, USA), Phospho-histone H2A.x (S139; Cell Signaling Technology), PrxI, PrxII, PrxIII, and Prx-SO₂/₃ (all from Abclone, Seoul, Korea), Srx, cIAP-1, and Tubulin (all from Santa Cruz Biotechnology, Dallas, TX, USA), and CD133 (including CD133 antibodies conjugated to magnetic beads and APC; Miltenyi Biotec, Auburn, CA, USA). The Srx inhibitor frugoside was isolated from Cardiospermum halicacadum, as previously described [20 ].

Four human colorectal cancer cell lines, HCT116, KM12C, HT29, and SW480, were purchased from the Korean Cell Line Bank (Korean Cell Line Research Foundation, Seoul, Korea) and maintained in RPMI1640 supplemented with 10% fetal bovine serum (Gibco BRL/Life Technologies, Inc., Gaithersburg, MD, USA). They were cultured at 37 °C in a humidified chamber containing 5% CO₂. Human recombinant TRAIL purchased from Sigma were used at indicated concentrations.