Figure 3 | Cell Death & Disease

Figure 3

From: MiRNA199a-3p suppresses tumor growth, migration, invasion and angiogenesis in hepatocellular carcinoma by targeting VEGFA, VEGFR1, VEGFR2, HGF and MMP2

Figure 3

VEGFA in cancer cells and VEGFR1 and VEGFR2 on endothelial cells are suppressed by miR-199a-3p. (a) Luciferase-reporter assay with VEGFR2 were determined separately in premiR-199a-3p-transfected wild-type and miR-199a-3p binding-site mutant 3′-UTRs of VEGFA, VEGFR1, HepG2 cells. (b) Ectopic expression of miR-199a-3p was confirmed at different time points. (c) Protein expression of VEGFA and VEGFR2 in SNU449 and HUVEC cells, respectively. By ELISA, secreted VEGFA was determined in (d) culture media of control vector, premiR-199a-3p and premiR-199a-3p with anti-miR-199a-3p-transfected SNU449 cells and (e) serum of healthy volunteer (n=8) and HCC patients (n=7). (f) Tube-formation assay was performed with premiR-199a-3p or vector-transfected HUVECs in the presence of 20 ng/ml of recombinant VEGFA and anti-VEGF antibody or control IgG-treated CM of SNU449. (g) VEGFR2, ERK1/2 and phospho-ERK1/2 levels were determined in transfected HUVEC cells cultured in the presence of 20 ng/ml of recombinant VEGFA. α-Tubulin was used as loading control in western blot. *P<0.05, **P<0.01 and ***P<0.005

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