Ht 29 cells

HT-29 cells were purchased from Jiangsu KeyGEN BioTECH Corp. (Jiangsu, China). The cells were cultured at 37°C and 5% CO₂ in DMEM 1640 (Gibco) that contained 10% fetal bovine serum (FBS, Gibco) and 1% penicillin–streptomycin (Gibco). When the cells reached 80% confluence, they were divided and sub-cultured for further passages with 0.25% trypsin and 0.02% ethylene-diamine-tetra-acetic acid (EDTA, Gibco). Cell viability was determined using the cell counting kit-8 (CCK-8) assay (APExBIO Technology LLC, United States). Briefly, cells were seeded and treated with different concentrations of luteolin (LUT, Aladdin, Figure 2A), β-sitosterol (SIT, Aladdin, Figure 2B), myristic acid (MYA, J&K Scientific, Figure 2C), and vanillin (VAN, J&K Scientific, Figure 2D) for 24 h, and 10% CCK-8 solution was added to each well. After incubation for 2 h at 37°C, the absorbance at 490 nm was read on a microplate reader (iMark, Bio-Rad, United States).

HCT116 cells, RKO cells, SW480 cells, HCT8 cells, HT29 cells, LOVO cells, and FHC cells were obtained from the KeyGEN BioTECH (Nanjing, Jiangsu, China). Cell culture was performed in an atmosphere of 5% CO₂ at 37°C.