Anti mir 497 5p

Four CRC cell lines (including LOVO, SW480, HT-29, and HCT-116) and one normal colon epithelial cell line (FHC) were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). All cell lines were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Tianhang, Hangzhou, China) and 1% penicillin-streptomycin in a humidi ed incubator (37°C, 5% CO 2 ). Small interfering RNA targeting XIST (si-XIST: 5'-GCCCUUCUCUUCGAACUGUTT-3') and its matching control (si-NC: 5'-CGTTAATCGCGTATAATACGCGTAT-3'), were obtained from Genepharma (Shanghai, China). A miR-497-5p mimic (miR-497-5p), its inhibitor (anti-miR-497-5p), and their corresponding controls (miR-NC and anti-miR-NC) were obtained from Ribobio (Guangzhou, China). For overexpression vectors, the sequence of FOXK1 was cloned into the pcDNA3.1 vector (vector) according to the manufacturer's instructions (Life Technologies). Brie y, 100 nM of miR-497-5p mimics or miR-497-5p inhibitor and 1000 ng plasmid were transfected into each 6-well plate for 48 h using Lipofectamine 3000 (Invitrogen, Waltham, MA, USA). For animal experiments, cells were stably transfected with an XIST lentiviral vector, constructed by Hanyin Biotechnology Co., Ltd., Shanghai, China. After transfection, DMEM medium was replaced with DMEM supplemented with puromycin (3 µg/mL) as a positive selection for infected cells.

Four CRC cell lines (including LOVO, SW480, HT-29, and HCT-116) and one normal colon epithelial cell line (FHC) were obtained from Shanghai Institute of Biochemistry and Cell Biology (Shanghai, China). All cell lines were cultured in DMEM (Gibco, USA) supplemented with 10% fetal bovine serum (FBS; Tianhang, Hangzhou, China) and 1% penicillin-streptomycin in a humidi ed incubator (37°C, 5% CO 2 ). Small interfering RNA targeting XIST (si-XIST: 5'-GCCCUUCUCUUCGAACUGUTT-3') and its matching control (si-NC: 5'-CGTTAATCGCGTATAATACGCGTAT-3'), were obtained from Genepharma (Shanghai, China). A miR-497-5p mimic (miR-497-5p), its inhibitor (anti-miR-497-5p), and their corresponding controls (miR-NC and anti-miR-NC) were obtained from Ribobio (Guangzhou, China). For overexpression vectors, the sequence of FOXK1 was cloned into the pcDNA3.1 vector (vector) according to the manufacturer's instructions (Life Technologies). Brie y, 100 nM of miR-497-5p mimics or miR-497-5p inhibitor and 1000 ng plasmid were transfected into each 6-well plate for 48 h using Lipofectamine 3000 (Invitrogen, Waltham, MA, USA). For animal experiments, cells were stably transfected with an XIST lentiviral vector, constructed by Hanyin Biotechnology Co., Ltd., Shanghai, China. After transfection, DMEM medium was replaced with DMEM supplemented with puromycin (3 µg/mL) as a positive selection for infected cells.