Figure 1. Plumbagin reduced the migration and invasion of the human endothelial cell line EA.hy926 that was induced by the human hepatoma cell line Hep3B cells.

(A) Plumbagin-depleted cells (24 h) were loaded for transwell migration (left) and Matrigel invasion assays (right). (B–C) Migration or invasion were assessed at 24 h. Fields were counted for each well. The Hep3B cells were treated with plumbagin as indicated and migration or invasion experiments were performed as in (B–C). (D) The co-cultured hy926 cells can also spontaneously form capillary-like structures on Matrigel, and we therefore studied the effects of plumbagin on the angiogenesis in hy926 cells. Our data showed that the number and the continuity of the capillary-like structures of the hy926 cells were all dramatically inhibited by 1.25–5 μM plumbagin in a dose-dependent manner, which suggested that plumbagin inhibited the formation of tubes that was induced by the hy926 cells in vitro. (E) The FITC–phalloidin staining assay was performed to examine the structure of F-actin. Plumbagin (5 μM) suppressed the changes in cell morphology and actin remodeling induced by the co-culture with hy926 cells. Thalidomide (25 μM) was used as the positive control, GSK690693 (10 μM) was used as the Akt inhibitor group. The data represent the mean values of three experiments ± SE.*P < 0.05, **P < 0.01, ***p < 0.001 compared to co-culture with hy926 cells.