Research Paper Volume 15, Issue 16 pp 8367—8383

Figure 6. circFAM169A enhances CRC angiogenesis and metastasis by targeting angiopoietin-2 (ANGPT2). (A) Venn diagram of mRNA that was highly correlated with circFAM169A and the downstream target genes of miR-518a-5p as predicted by TargetScan. (B) Survival analysis of ANGPT2. (C) ANGPT2 expression in normal and tumor tissues. The luciferase activities were measured by a dual-luciferase assay, and the Renilla/firefly luciferase light-unit ratio was measured. (D) ROC curve showed the diagnostic value of ANGPT2. (E) Construction of a luciferase plasmid encoding mut-ANGPT2. (F) The luciferase activities of different groups were measured by a dual-luciferase assay. (G, H) Transwell migration assay and Transwell invasion assay were performed using HCT116 and RKO cells. All data are presented as means ± SD (n = 3 independent experiments). ^***p ≤ 0.001 and ^****p ≤ 0.0001. (I) Western blotting analysis of ANGPT2, total TIE2, and p-TIE2 expression in circFAM169A-knocked down HCT116 cells treated with miR-518a-5p mimics. (J) Western blotting analysis of ANGPT2, total TIE2, and p-TIE2 expression in circFAM169A-overexpressing RKO cells treated with miR-518a-5p inhibitors. (K) Western blotting analyses of total FAK, p-FAK, total AKT, and p-AKT in CRC cells with different circFAM169A expression levels.