Fig. 4.

The cell proliferation, migration, and angiogenic abilities of endothelial cells of HCC are suppressed after silencing of HepG2-secreted BMP2. To evaluate the modulatory role of BMP2 in angiogenesis, the HepG2 cells infected with lentivirus expressing oeBMP2 or shBMP2 were co-cultured with ECV304 cells, and VEGF was delivered to HepG2 cells to explore how it functions in a co-culture system. a Proliferation of ECV304 cells assessed by MTT assay. b VEGF expression patterns in the supernatant of medium evaluated by ELISA. c Migration of ECV304 cells tested by Transwell assay (× 400). d Angiogenic ability assessed by tube formation assay (× 200). e Western blot analysis for expression patterns of angiogenesis-related (VEGF, VEGF-C) and invasion-related (MMP-2, MMP-9, Vimentin, and E-cadherin) proteins. Data were expressed as mean ± standard deviation, and data between two groups were compared by Student’s t test. *p < 0.05 compared with the NC group. HCC, hepatocellular carcinoma; BMP2, bone morphogenetic proteins 2; oe, over-expression NC, negative control; VEGF, vascular endothelial growth factor; MTT, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H_tetrazolium bromide; ELISA, enzyme-linked immunosorbent assay