Figure 6
Identification of the key targets and pathways of VS-6063 and JQ1 in ovarian tumor cells. (a) Expression of total or phosphorylated FAK or Myc in human ovarian tumor cell lines. Blotting for lanes 1–8 and lanes 9–12 was conducted separately with the lysates from the indicated human HGSOC cell lines. (b) OVC-8 or OVC-433 cell lines were treated with indicated inhibitors for 48 h, followed by analyses of levels of total or phosphorylated FAK or Myc proteins. Equal protein loading for OVC-8 cells was also assessed by blotting β-actin shown in Figure 5c. (c) OVC-8 cells were treated with siRNA for 24 h, and followed by incubation with 2 μm VS-6063 for additional 48 h. Tumor cells were then analyzed by the MTT assay or blotted for c-Myc. Values (right panels): mean±s.e., n=3. *P-value <0.05. (d) Effect of VS-6063 and JQ1 on key downstream pathways of the integrin–FAK signaling axis. OVC-8, OVC-433 and SK-OV-3 cells were treated with different combinations or dosages of VS-6063 and JQ1 alone or in combination for 48 h and analyzed for the activation of the PI3k/AKT or Ras/MAPK pathway. (e) Effect on the cell survival-related pathway. Tumor cells treated with various doses of inhibitors for 48 h and analyzed for differences in cleaved Caspase 8, XIAP and Bim by immunoblotting. β-Actin was blotted as a control for OVC-433 or OVC-8 (shown in Figure 5c).
