Colo201

The colon cancer cell lines (SW620, LoVo, SW480, HCT116, SW48, HCT15, RKO, SW837, COLO-201, COLO-205, LS174T and LS180) used in the present study were originally obtained from the American Type Culture Collection (Manassas, VA, USA). The cells were cultured in the recommended media (Dulbecco's modified Eagle's medium (Thermo Fisher Scientific, Inc., Waltham, MA, USA) for RKO, LS174T and LS180 and RPMI-1640 medium (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) for SW480, Lovo, HCT15, SW48, SW620 and HCT116 supplemented with 10% fetal bovine serum (Thermo Fisher Scientific, Inc.). The monolayer cells were maintained in a 37°C incubator with 5% CO₂, observed regularly under a light microscope (magnification, ×40) and subcultured when they reached 80–90% confluency.

Four human colorectal cancer cell lines were used in the present study. HT29, colo 201, HCT116, HCT15, and CCD-18Co cells were purchased from the American Type Culture Collection (Manassas, VA, USA). HT29 and HCT116 cells were maintained in McCoy's 5A (Gibco, Grand Island, NY, USA) medium containing 10% fetal bovine serum (FBS) (Gibco), 100 U ml⁻¹ of penicillin G, and 100 μg ml⁻¹ of streptomycin at 37 °C in a humidified 5% CO₂ atmosphere. HCT-15 cells were maintained in Eagle's minimum essential medium (Sigma-Aldrich, St Louis, MO, USA) containing 10% FBS (Gibco), 100 U ml⁻¹ of penicillin G, and 100 μg ml⁻¹ of streptomycin at 37 °C in a humidified 5% CO₂ atmosphere. colo 201 cells were maintained in Roswell Park Memorial Institute 1640 Medium (Sigma-Aldrich) containing 10% FBS (Gibco), 100 U ml⁻¹ of penicillin G, and 100 μg ml⁻¹ of streptomycin at 37 °C in a humidified 5% CO₂ atmosphere. CCD-18Co cells were maintained in Eagle's minimum essential medium (Sigma-Aldrich) containing 10% FBS, 100 U ml⁻¹ of penicillin G, and 100 μg ml⁻¹ of streptomycin at 37 °C in a humidified 5% CO₂ atmosphere.

The colon cancer cell lines (HCT116, LoVo, RKO, LS174T, Colo205, Colo201, SW620, LS180, SW837, SW480, HCT15, and SW48) used in the present study were originally obtained from the American Type Culture Collection (Manassas, VA, USA). RKO, LS174T and LS180 cells were cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (both from Thermo Fisher Scientific, Inc., Waltham, MA, USA). SW480, LoVo, HCT15, SW48, SW620 and HCT116 cells were cultured in RPMI-1640 (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany) supplemented with 10% fetal bovine serum. The monolayer cells were maintained in a 37°C incubator with 5% CO₂, observed regularly under a light microscope (×40) and subcultured when 80–90% confluence was reached.

Human epidermoid carcinoma A431 cells, human esophageal cancer EC17 cells and human prostate cancer PC‐3 cells were provided by M. Kawada (Institute of Microbial Chemistry, Japan). Human esophageal cancer EC109 cells were provided by Columbia University (New York, NY, USA). Human cervical cancer HeLa cells were provided by M. Yoshida (RIKEN, Japan). Human embryonic kidney HEK293T cells were provided by S. Saiki (Juntendo University, Japan). Human colorectal tumor LoVo, HT29, Colo‐201, HCT116, LS‐174T, SW620, DLD‐1, SW48 and SW480 cells, human lung cancer A549 cells, human breast cancer MCF‐7 cells, and human melanoma A2058 cells were obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA).
A431 cells were maintained in DMEM supplemented with 5% calf serum (CS), 100 U/mL penicillin G (Sigma‐Aldrich, St. Louis, MO, USA), and 0.1 mg/mL kanamycin (Sigma‐Aldrich) at 37°C in a humidified atmosphere containing 5% CO₂. EC17, HEK293T, HeLa and MCF‐7 cells were maintained in DMEM supplemented with 10% FBS, 100 U/mL penicillin G, and 0.1 mg/mL kanamycin in the same conditions described above. The other cells were maintained in RPMI‐1640 medium supplemented with 10% FBS, 100 U/mL penicillin G and 0.1 mg/mL kanamycin, also in the abovementioned conditions.

RH7777 rat hepatoma cells were kindly donated by Dr Chiba K (Mitsubishi Tanabe Pharma). Human CRC cell lines (Caco2, COLO 201, COLO 205, COLO 320DM, HCT116, HT29, LS1034, LS123, LS174T, LS180, RKO, RKO‐E6, SNU‐C1, SW1116, WiDr), PK‐1, PK‐59, PANC‐1, MIA PaCa‐2 pancreatic, BT20, BT474 breast, A549, NCI‐H2170 lung cancers, and P3U1 mouse myeloma cells were purchased from the American Type Culture Collection (ATCC). CCK‐81 and OUMS23 human CRC cells were purchased from JCRN Cell Bank. SW‐C4 is a clone originating from SW1116, and LS‐LM4¹¹ is a highly liver‐metastatic clone from LS174T. The HEK293F human embryonic kidney cell line was purchased from Invitrogen. All cells were cultured in RD medium,¹² which is a blended medium of equivalent volumes of RPMI‐1640 and Dulbecco's modified Eagle medium (Nissui Pharmaceutical Co, Ltd) containing 7% heat‐inactivated fetal bovine serum (FBS, Thermo Fisher Scientific Inc) at 37°C under humid conditions. Aseptic processing during cell culture was ideally controlled by a MediAir air purifier (Pieras Co., Ltd) equipped in the cell‐culture room. Cell growth was measured as previously described.¹³ Briefly, a WST‐8–based cell counting kit (Dojin Chemicals) was used, and 450 nm was measured using a microplate reader.