Human leukemic cell line K562 cells were purchased from ATCC (Manassas, VA, USA), and cultured in RMPI 1640 medium, which is supplemented with 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin in a humidified 5% CO2 incubator at 37°C. Imatinib was purchased from Sigma (St. Louis, MO, USA), and Imatinib-resistant K562 cells were constructed following the method previous described (Kang et al., 2014 (link)). K562 cells were transfected with wild type or mutant MEG3 construct using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer’s instructions. MicroRNA-21 (miR-21) mimics or mimic control (GenePharma, Shanghai, China) at a final concentration of 25 nmol/L was transfected into Imatinib-resistant K562 cells using Lipofectamine 2000 (Invitrogen) according to the manufacturer’s instructions.
Protocol detail
Imatinib-resistant K562 Cell Line
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Cell line
Cultured medium
Fetal bovine serum
Human
Imatinib
K562 cells
Lipofectamine 2000
Microrna
Penicillin
Streptomycin
Corresponding Organization : Affiliated Hospital of Southwest Medical University
Other organizations : Nanchong Central Hospital, The University of Texas MD Anderson Cancer Center
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Variable analysis
independent variables
- Transfection of wild type or mutant MEG3 construct
- Transfection of microRNA-21 (miR-21) mimics or mimic control
- Not explicitly mentioned
- K562 cells were cultured in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 100 U/ml penicillin, and 100 μg/ml streptomycin in a humidified 5% CO2 incubator at 37°C
- Imatinib-resistant K562 cells were constructed following the method previously described (Kang et al., 2014)
- Transfection of microRNA-21 (miR-21) mimics
- Transfection of mimic control
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