Fig. 5: ZFAS1 functioned as a competing endogenous RNA (ceRNA) by sponging miR-150-5p.

a The expression of miR-150-5p and miR-590–3p were detected in HCT116 and HCT8 cells after transfecting with pcDNA3.1-ZFAS1 or blank vector. b The expression of miR-150-5p was detected using qRT-PCR after the biotinylated-ZFAS1 pull down assay in HCT116 cells. c The biotinylated wild-type/mutant miR-150-5p was transfected into HCT116 cells with ZFAS1 overexpression. The expression levels of ZFAS1 were measured by qRT-PCR after streptavidin capture. d Wild and mutant ZFAS1 sequences were cloned into pmirGLO reporter, Luciferase activity in HCT116 and 293T cells cotransfected with agomiR-150-5p or agomiR-NC and pmirGLO-ZFAS1-WT or pmirGLO-ZFAS1-Mut. Luciferase activities were normalized to renilla luciferase. e Anti-AGO2 RIP was used in HCT116 cells overexpressing agomiR-150-5p, followed by qRT-PCR to evaluate the expression of ZFAS1 or H19 (control) associated with AGO2. The data are shown as the mean ± SD of three independent experiments. f miR-15-5p was downregulated in CRC tissues compared to paired adjacent normal tissues. g The expression of miR-150-5p in HCT116, HCT8, HT29, SW620, SW480, DLD-1, and FHC. h The correlation between ZFAS1 level and miR-150-5p level in CRC tissues. i CCK-8 proliferation assays in siZFAS1-1 and antagomiR-150-5p transfected HCT116 and HCT8 cells. j Wound healing assays in siZFAS1-1 and antagomiR-150-5p transfected HCT116 and HCT8 cells. k Transwell invasion assays in siZFAS1-1 and antagomiR-150-5p transfected HCT116 and HCT8 cells. l HUVECs tube formation in siZFAS1-1 and antagomiR-150-5p transfected HCT116 and HCT8 cells. Each experiment were performed three times. Data were shown as mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001