Normal colon FHC cells and four CRC cell lines (SW480, HCT116, SW620, LoVo) were purchased from COBIOER (Nanjing, China) and cultured in Dulbecco’s modified eagle medium (DMEM; Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Carlsbad, CA, USA), 100 U/mL penicillin and 100 μg/mL streptomycin at 37°C under a humidified atmosphere containing 5% CO2.
Short hairpin RNA (shRNA) targeting circ_0000512 (sh-circ#1, sh-circ#2 and sh-circ#3) and corresponding negative control (sh-NC), RUNX1 overexpression vector (RUNX1) and empty vector (vector) were purchased from Genechem (Shanghai, China). MiR-296-5p mimic or inhibitor (miR-296-5p or anti-miR-296-5p) and their negative controls (miR-NC or anti-NC) were synthesized by GenePharma (Shanghai, China). HCT116 and SW620 cells were transfected with these oligonucleotides (50 nM) or vectors (2 μg) by Lipofectamine 3000 reagent (Invitrogen).
Short hairpin RNA (shRNA) targeting circ_0000512 (sh-circ#1, sh-circ#2 and sh-circ#3) and corresponding negative control (sh-NC), RUNX1 overexpression vector (RUNX1) and empty vector (vector) were purchased from Genechem (Shanghai, China). MiR-296-5p mimic or inhibitor (miR-296-5p or anti-miR-296-5p) and their negative controls (miR-NC or anti-NC) were synthesized by GenePharma (Shanghai, China). HCT116 and SW620 cells were transfected with these oligonucleotides (50 nM) or vectors (2 μg) by Lipofectamine 3000 reagent (Invitrogen).