Kyse270

Human ESCC cell lines (KYSE140, KYSE150, and KYSE270) were purchased from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). KYSE30 was obtained from HPA Culture Collections. These cell lines were maintained in Ham's F12/RPMI1640 medium containing 2% FBS. Human esophageal epithelial cells (HEEpiC) were purchased from ScienCell Research Laboratories, Inc. (Carlsbad, CA) and were maintained in Epithelial Cell Medium-2 (ScienCell Research Laboratories). In some experiments, the cells were cultured in a 24-well plate at a density of 5 × 10⁴ cells/well for 18 h, and then treated with 5-aza-2′-deoxycytidine (5-aza-dC; Sigma-Aldrich, Inc., St. Louis, MO) for 72 h.

Fresh tumor tissues were obtained from ESCC patients. Samples were diced, digested with digestive juice containing 1 mg/mL collagenase type I (Sigma-Aldrich, S.t. Louis, MO, USA), 1 mg/mL collagenase type IV (Sigma-Aldrich), filtered, then resuspended in Dulbecco’s Modified Eagle Medium (DMEM)/F-12 Medium (Gibco, Carlsbad, CA, USA), supplemented with 5% Fetal Bovine Serum (FBS, Gibco). The culture medium was changed every 3 days. Fibroblasts were removed by incubation with 0.05%Trypsin/EDTA. The primary patient-derived tumor cells were named SBRC-EC01 and SBRC-EC02. For the experiments described here, all cells were used in passages 10 to 16. The cell lines KYSE510, KYSE180, KYSE270 and other ESCC cell lines were obtained from the Japanese Collection of Research Bioresources Cell Bank (JCRB, Osaka, Japan), and passaged in Roswell Park Memorial Institute-1640 (RPMI 1640, Gibco) with 10% FBS. The human kidney epithelial cell line HEK-293 was purchased from the Cell Bank of the Type Culture Collection Committee of the Chinese Academy of Sciences (Shanghai, China) and maintained in DMEM supplemented with 10% FBS. All cells were verified free of mycoplasma and bacterial contamination.

A total of 21 ESCC cell lines, specifically KYSE30, KYSE70, KYSE140, KYSE150, KYSE180, KYSE270, KYSE410, KYSE450, KYSE510, KYSE590, KYSE890, KYSE1170, KYSE1260, KYSE1440, NUEC2, TE1, TE2, TE3, WSSC, TT, and TTn, along with Het‐1A, a non‐cancerous epithelial cell line, were purchased from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan), the American Type Culture Collection (Manassas, VA, USA), or were established at Nagoya University.
⁵ (link) The cells were cultured in a humidified incubator at 37°C with an atmosphere composition of 5% CO₂ using RPMI‐1640 (Sigma‐Aldrich, St. Louis, MO, USA) supplemented with 10% fetal bovine serum and antibiotics.
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