Sw620

The human colorectal cancer cell line SW620 and LS180 were purchased from Cell Resource Center, Peking Union Medical College (Beijing, China). The ABCB1-overexpresison SW620/ADR cell line was selected as previously described [21 (link)]. Briefly, SW620 cells were grown in complete DMEM medium, stepwise exposure to doxorubicin, the beginning exposure concentration was 30 nM, which finally increased to 300 nM for 3 months. Finally, a clonal cell line that grew in the presence of 0.3 μM doxorubicin were isolated as SW620/ADR. The ABCG2-overexpression LS180/MX cell line was selected by increasingly exposure to mitoxantrone from 1 μM to 80 μM as previously described [22 (link)]. Finally, a clonal cell line that grew in the presence of 80 μM mitoxantrone was isolated as LS180/MX. HEK293/pcDNA3.1, HEK293/ABCB1, and HEK293/ABCG2 cells were transfected with pcDNA3.1 vector or vector with full length of ABCB1 or ABCG2, which were kindly obtained from Zhejiang University (Zhejiang, China). HEK293/ABCB1, and HEK293/ABCG2 cells were selected with complete medium containing of G418 (2 mg/ml). All of the cell lines were cultured in 10% FBS DMEM medium containing 5% CO₂ and humid atmosphere.

Human colorectal cancer cell lines (HCT-116 and SW620) were commercially purchased from the cell resource center of the Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences (Shanghai, China), and cultured in McCoy’s 5 A medium and Leibovitz’s L-15 medium supplemented with 10% fetal bovine serum and 1% penicillin-streptomycin, respectively, at 37 ℃ in a humidified incubator with 5% CO₂.

The human colorectal cancer cell lines SW-480 and SW-620 were purchased from the Cell Resource Center, IBMS, CAMS/PUMS and passed in less than 6 months. Cells were cultured in RPMI-1640 (Gibco, Paisley, UK) with 10% FBS (Gibco, Paisley, UK) and 2 mmol/L L-glutamine (Gibco, Paisley, UK), 100 U/ml of penicillin (Gibco, Paisley, UK), and 100 µg/ml of streptomycin sulfate (Gibco, Paisley, UK).

Human colon cancer SW620 cells were purchased from Cell Resource Center (IBMS, CAMS/PUMC, Beijing, China) characterized by STR Profiling. Cells were cultured in Dulbecco's modified Eagle's medium, supplemented with 10% fetal bovine serum, 100 units/ml penicillin, and 100 µg/ml streptomycin at 37 °C in 5% CO₂.

Human CRC cells HCT8 and SW620 were obtained from Cell Resource Center, Peking Union Medical College (Beijing, China). The SW620 cell line and its doxorubicin-selected ABCB1-overexpressing SW620/Ad300 cell line were a gift from Drs. Susan E. Bates and Robert W. Robey (National Cancer Institute (NCI), National Institutes of Health (NIH); Bethesda, MD, USA) and they were used for the ABCB1 reversal study. Human umbilical vein endothelial cells (HUVECs) were purchased from Cell Bank of Chinese Academy of Sciences (Shanghai, China). HCT8 and SW620 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% fetal bovine serum (FBS) and SW620/Ad300 cells were maintained in medium with 300 ng/mL doxorubicin. HUVECs were cultured in Dulbecco's Modified Eagle's Medium supplemented with 10% FBS. All the cells were cultured at 37°C in a humidified atmosphere containing 5% CO₂. Drug-resistant cells were grown in drug-free culture media for >2 weeks before assay.