Developing ATB-Resistant Cancer Cell Lines

HCT116 and LOVO cells (1 × 10⁷) were exposed to 100 mg/mL N-ethyl-N-nitrosourea (Sigma-Aldrich, Burlington, MA, USA) for 24 hours, followed by treatment with a gradient concentration of ATB to induce ATB resistance. During the first five days, HCT116 and LOVO cells were exposed to ATB (4 μg/mL), and the medium was changed every day. In the next two months, 6, 8, 10, and 12 μg/mL of ATB were used to treat the cells [32 (link)]. Finally, about 100 variable cells exposed to ATB (12 μg/mL) were recovered. After proliferation for about one month, the ATB-resistant HCT116 and LOVO cells, named HCT116/ATB and LOVO/ATB separately, were used for functional assays.