Figure 2

LBH589 decreases the levels of p-STAT3, p-Akt and Bcl-xL in HCC cells, and gankyrin overexpression can attenuate the effect of LBH589. LBH589 mediates the expression of p-Akt and p-STAT3 through gankyrin/PI3K/Akt and gankyrin/Rb/IL-6/JAK2/STAT3 pathways. (A) HCC cells were treated for 24 h with or without LBH589 and analyzed for the indicated protein by western blotting. (B) Western blotting results showed LBH589 induced Akt and STAT3 dephosphorylation was reduced by gankyrin overexpression. (C) Western blotting results showed LBH589 decreased Bcl-xL expression and overexpression of gankyrin partially protected against LBH589-induced inhibition of Bcl-xL. β-actin was used as the internal control. All assays were done in triplicate. (D) LBH589 increased Rb expression and decreased the expression of PI3K, p-PI3K (Tyr458), JAK2 and p-JAK2 (Tyr1007/1008) in three HCC cells. After treatment of LBH589, the expression of p53 increased in HepG2, no obvious change was detected in HCC-LM3 and SMMC-7721 cells. β-actin was used as the internal control. All assays were done in triplicate. (E) After treatment of LBH589, the level of IL-6 in three HCC cells was significantly decreased compared with controls. The results represent means ± SD of experiments performed in triplicate. ***P < 0.001, LBH589 treated versus control group.