Figure 1

LBH589 is cytotoxic to HCC cells in a dose- and time-dependent manner. (A) MTT assay showing percentage of viable HCC cells treated with DMSO, 1, 5, 10, 50, 100 and 1000 nM of LBH589 for 24, 48 and 72 h. The results represent means ± SD of experiments performed in triplicate. ***P < 0.001, LBH589 treated versus control group. (B) Expression of acetyl-Histone H3, acetyl-Histone H3 (Lys27) and acetyl-Histone H4 were determined via western blotting after treatment with 50 and 100 nmol/L LBH589 for 24 h. Tubulin was used as the internal control. All assays were done in triplicate. (C) The expression of gankyrin after treatment with 50 and 100 nmol/L LBH589 for 24 h. β-actin was used as the internal control. All assays were done in triplicate. (D) Flow cytometry results of annexin V-PI staining of HCC cells after exposure to DMSO or LBH589 (50 nM) for 48 h. The results represent means ± SD of experiments performed in triplicate. ***P < 0.001, LBH589 treated versus DMSO group. (E) A representative example of apoptosis of HepG2 cell line treated with 50 nM of LBH589 at 48 h is shown. (F) Expression of apoptosis relative proteins were determined via western blotting after treatment with 50 and 100 nmol/L LBH589 for 24 h. β-actin was used as the internal control. All assays were done in triplicate. (G) Flow cytometry results showed gankyrin overexpression attenuated the LBH589-induced apoptosis of HCC cells. The results represent means ± SD of experiments performed in triplicate. ***P < 0.001, LBH589 + vector ctrl treated versus LBH589 + gankyrin group. (H) A representative example of apoptosis of HepG2 cell line (which transfected human gankyrin/control plasmid) treated with 50 nM of LBH589 at 48 h.