Hct116

SW480 and HCT116 cells were obtained from China Infrastructure of Cell Line Resources (Beijing, China). Cell line authentication and mycoplasma testing were not performed. SW480 was cultured in Dulbecco's Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS). HCT116 was cultured in Iscove's modified Dulbecco's medium (IMDM) containing 10% FBS. All cell cultures were maintained at 37 °C under at 5% CO₂ in a humidified atmosphere. Cells were passaged approximately every 2–3 days.

MCF-7 and HCT116 from the China Infrastructure of Cell Line Resources were cultured in DMEM and IMDM (Gibco), respectively. H1299 from the American Type Culture Collection (ATCC) was cultured in RMPI-1640 culture medium (Gibco). All cell culture media contained 10% PBS, 100 U ml⁻¹ penicillin, and 100 μg ml⁻¹ streptomycin. Cells in exponential growth were seeded at 2000–3000 cells per well in 96-well culture plates and allowed to grow for 24 h before treatment. Culture media were added to stock molecular solutions and final molecular formulations contained 0.2% DMSO (v/v) and 0.01% Tween 80 (v/v). Spectroscopic reading was taken 72 h after treatment, and the cell survival percentage was calculated as the percentage absorbance of treated wells versus reference wells.

Human colon cancer cell lines LoVo and SW620 were purchased from the Cell Bank of the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China). All LoVo, SW620 and HCT116 (China Infrastructure of Cell Line Resources, Beijing, China) cells were grown in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) and 1% penicillin and streptomycin. The cell lines were maintained in a humidified incubator at 37°C with an atmosphere of 5% CO₂.

Various cell lines (HCT116, DLD1, MCF7, LS174T, MDA-MB-435, and MIApaca-2) were obtained from the China Infrastructure of Cell Line Resources (Kunming, Beijing, or Shanghai, China). HEK-293T cells were obtained from A. Lasorella (The Institute for Cancer Genetics, Columbia University Medical Center, New York, NY). HCT116, DLD1, MCF7, MDA-MB-435, MIApaca-2 and HEK-293T cells were cultured in DMEM (Life Technologies) supplemented with 10% FBS (Life Technologies), 100 U/mL penicillin (Life Technologies), and 100 mg/mL streptomycin sulfate (Life Technologies). LS174T cells were cultured in RPMI 1640 (Life Technologies) supplemented with 10% FBS, 100 U/mL penicillin, and 100 mg/mL streptomycin sulfate. LS174T suspended cell spheres, named LS174T-CSC, were established using serum-free neural stem cell medium composed of DMEM/F12, EGF (20 ng/ml, Life Technologies, USA), bFGF (20 ng/ml, Life Technologies, USA), and B27 (1x, Life Technologies, USA). All the cells were cultured at 37°C in a humidified incubator with 5% CO2 and routinely confirmed to be mycoplasma free by PCR. The cells were passaged when they reached approximately 80% confluency.