Figure 4.

MiR-590-5p inhibits VEGFA expression and CRC angiogenesis by repressing NF90. (a) Schematic representation of the putative miR-590-5p target site within the 3'-UTR of NF90 was shown on the top. Mutations used for abolishing miR-590-5p recognition sequence were highlighted in red. (Bottom) SW620 cells were transfected with control or pre-miR-590 plasmid along with WT or mutant NF90 luciferase reporter as shown. The activity of both Renilla and Firefly luciferases was assayed using the dual-luciferase reporter assay system. (b and c) Indicated cells were, respectively, transfected with pre-miR-590 or miR-590-5p inhibitor or their control plasmids. α-tubulin was used as loading control, and the levels of NF90 mRNA and protein expression were determined by qRT-PCR and western blot, respectively. (d) NF90 expression was determined by western blot in HCT116 Dicer+/+ and Dicer−/− cells transfected with control or pre-miR-590. (e and f) Western blotting assay was used to analyze the protein levels of VEGF and NF90 in indicated CRC cells. (g) NF90 expression was determined by western blot in CRC and adjacent tissues. (h) Linear correlation analysis was used to detect the correlationship between the levels of miR-590-5p and NF90 expression. *P<0.05, **P<0.01