Glycer-AGEs-RAGE signaling enhances the angiogenic potential of hepatocellular carcinoma by upregulating VEGF expression

Figure 3 Effect of glyceraldehyde-derived advanced glycation end-products on the malignancy of hepatocellular carcinoma cells. Hep3B and HepG2 cells were incubated with control unglycated bovine serum albumin (BSA) or glyceraldehyde-derived advanced glycation end-products (Glycer-AGEs) for 24 h. A: In Hep3B cells, cyclooxygenase-2 (COX-2) mRNA expression levels were analyzed using real-time reverse transcription-polymerase chain reactions, and results were normalized to the β-actin mRNA level (n = 3), ^bP < 0.01 vs control unglycated BSA; B: COX-2 expression as measured by Western blotting. Cell lysates (30 μg of proteins/lane) were loaded onto a 10% polyacrylamide gel. Size markers (kDa) are shown on the left. Equal protein loading was verified using an anti-β-actin antibody. As a positive control, A549 cells were incubated with phorbol 12-myristate 13-acetate (PMA: 100 nmol/L) for 6 h; C: The migratory capacity of Hep3B cells was evaluated using the Oris cell migration assay (n = 8). Cells were incubated with control unglycated BSA or Glycer-AGEs for 24 h, and the number of migrating cells was then assessed using a fluorescence microplate reader. RFU: Relative fluorescence units. The open and filled bars represent results for cells treated with control unglycated BSA and Glycer-AGEs, respectively. Data are shown as the mean ± SD.