Hct116 cells

The cell lines used were as follows: NCM460 cells (cat. no. CC-Y1550; EK-Bioscience GmbH; a human colon epithelial cell line), RKO cells (cat. no. CL- 196; Procell Life Science & Technology Co., Ltd.), HCT116 cells (cat. no. CL-0096, Procell Life Science & Technology Co., Ltd.) and SW620 cells (cat. no. CL-0225; Procell Life Science & Technology Co., Ltd.). RKO, HCT116 and SW620 are human COAD cell lines. Cells were cultured at 5% CO₂ and 37°C with DMEM (Sigma-Aldrich; Merck KGaA), as described in a previous study [14 (link)]. Cells were treated with 10 μg SKL2001 (Selleck Chemicals) for 7 days at 37°C for the last part of the experiments in vitro.

The human colorectal cancer HCT116 cells were obtained from Wuhan Procell Life Science & Technology Co., Ltd. (cat. no. CL‑0096). HCT116 cells were cultured in RPMI‑1640 medium (cat. no. AG29714278, Hyclone, Cytiva) supplemented with 10% FBS (cat. no. 11011‑8611, Zhejiang Tianhang Biotechnology Co., Ltd.) and 1% penicillin–streptomycin ((cat. no. ST488, Beyotime Institute of Biotechnology) in a 5% CO₂ incubator at 37 ℃. When the cells reached 80–90% confluence, the cells were digested with 0.05% trypsin cell digestion solution (cat. no. C0202, Beyotime Institute of Biotechnology) at 37 °C for about 1 min, and the subcultures were continued.