Hct 8 5 fu

The CRC cell lines SW620, SW480, HCT-8, HCT-8/5-FU and LoVo were purchased from Keygen Biotech Co. Ltd. (Nanjing, China). SW620 and SW480 were cultured in Leibovitz’s L-15 (Gibco, Grand Island, NY) medium contained 10% inactivated fetal bovine serum (Gibco, Grand Island, NY). HCT-8, HCT-8/5-FU and LoVo cells were cultured in in RPMI 1640 (Gibco, Grand Island, NY) medium contained 10% inactivated fetal bovine serum. To develop the 5-FU resistant LoVo cells, 5-FU was added to the medium by stepwise increasing concentrations for over 6 months. HCT-8/5-FU and LoVo/5-FU were maintained in medium supplemented with 124.5 μM and 113.0 μM 5-FU. CRC cells were both incubated at 37 °C with a humidified atmosphere containing 5% CO₂. All cells lines were routinely tested for mycoplasma, which were shown to be negative.

The human ovarian cancer cell line A2780 was purchased from KeyGen Biotech (China). The human liver cancer cell line HepG2, the human breast cancer cell line MDA-MB-231, the human colorectal cancer cell lines HCT-8 and LoVo along with their 5-FU-resistant strains (HCT-8/5-FU and LoVo/5-FU), and the human colon epithelial cell line HCoEpiC were purchased from the American Type Culture Collection (ATCC). A2780, LoVo/5-FU, HCT-8, HCT-8/5-FU, and HCoEpiC cells were cultured in RPMI 1640 medium (Gibco, New Zealand). LoVo cells were cultured in F-12K medium (Thermo). HepG2 cells were cultured in minimal essential medium (MEM) (Thermo). MDA-MB-231 cells were cultured in Leibovitz’s L-15 medium (Thermo). All culture media contained 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S). All cell lines were cultured in a humidified 5% CO₂ atmosphere at 37°C, except for MDA-MB-231 cells, which were cultured in a humidified 100% air atmosphere at 37°C. All tested RNA samples were dissolved in nuclease-free water and stored at −80°C before use. 5-FU was dissolved in dimethyl sulfoxide (DMSO) and used as a positive control.