Antisense oligonucleotide targeting midkine suppresses in vivo angiogenesis

Figure 1 RT-PCR (A) and Western-blotting analysis (B) of MK expression in human umbilical vein endothelial cells (HUVEC) after transfection with 0. 4 μmol/L MK-AS (lane 4), MK-SEN (lane 3), Lipofectin alone (lane 2) for 24 h. Lane 1 represents cells without transfection. GAPDH and β-actin were used as control respectively.

Figure 2 Effect of MK on growth of HUVEC. Cells after transfection with 0.4 μmol/L MK-AS or MK-SEN for 24 h were analyzed by MTS assay. Data were expressed as mean ± SD from four independent experiments.

Figure 3 Angiogenesis in normal chick CAM (A), HepG2-induced CAM (B), and HepG2-induced CAM treated with 0. 4 μmol/L MK-AS (C). Chick CAM assays were used to assess the impact of MK-AS on angiogenesis in vivo.

Figure 4 Immunohistochemical analysis of HCC xenograft angiogenesis after treatment with MK-SEN (A), 25 mg/kg of MK-AS per day (B), 50 mg/kg of MK-AS per day (C), and 100 mg/kg of MK-AS per day (D). Representative views of anti-CD34 antibody-stained vascular endothelium (brown) (х 200).

Figure 5 Tumors resected from mice at the experimental end point. Resected tumors were sectioned and stained as described in the text. Microvessel densities (MVD) were determined by counting CD34-positive endothelial cells in the sections and presented as mean ± SE positive cells/field from the three “hot-spot” fields. ^aP < 0.05, ^bP < 0.01 vs control group.