Ncm460

Three CRC cell lines, SW480, SW620, and HCT116, were purchased ATCC (Manassas, USA). A normal human colon mucosal epithelial cell line, NCM460, was obtained from Fenghui Biotechnology (Hunan, China). These cells were cultured in DMEM medium (Gibco, Carlsbad, CA, USA) containing 10% FBS (Absin, Shanghai, China) and 1% penicillin/streptomycin (ThermoFisher Scientific, Waltham, MA, USA), in a humidified atmosphere with 5% CO₂ at 37 °C.

Human colon cancer cell lines (HCT116 and DLD1) were purchased from Procell Company (Wuhan, China) and human colon mucosal epithelial cell line (NCM460) were purchased from Fenghui Biotechnology (Hunan, China). All cells were cultured in DMEM containing 10% fetal bovine serum (FBS) and 100 U/mL mixture of penicillin and streptomycin. The TDA is provided by the School of Pharmacy, Chongqing University. Cell counting kit 8 was purchased from MCE. Annexin V-FITC/PI Apoptosis Detection kit was purchased from CWBIO (Beijing, China). Primary antibodies for Western blotting, immunofluorescence and immunohistochemistry against target proteins are shown as follows: Caspase9/P35/P10 (Proteintech, Chicago, IL, USA, 66169), Caspase3 (Zen Bioscience, Chengdu, China, 300968), cleaved-Caspase3 (Zen Bioscience, 380169), PARP1 (Zen Bioscience, 380451), LC3 (Sangon Biotech, Shanghai, China, D163557), ATG5 (Sangon Biotech, D121650),SQSTM1/p62 (Beyotime, Shanghai, China, AF5312), β-actin (Santa Cruz, sc-47778), cyclinD1 (Cell Signaling Technology, USA, 2978S), cyclinB1 (Cell Signaling Technology, 4135S), PERK (Cell Signaling Technology, 3192S), IRE1 (Zen Bioscience, 220399), XBP1(Zen Bioscience, 381710), CHOP (Zen Bioscience, 381679) and HSPA5 (Sangon Biotech, D260466).