Figure 3

Gemcitabine attenuates Cisplatin-induced survival Akt signals and enhances apoptosis in Cisplatin-resistant cell lines. Caov-3 (A), A2780CP (B) and A2780 (C) cells were treated with or without 100 μM of Cisplatin for 24 hours in combination with 100 nM of Gemcitabine for 12 hours. The cell lysates were subjected to SDS-PA GE, followed by a Western blot analysis with an anti-phospho-Akt (Ser473) antibody (top parts of A, B and C) and an anti-Akt antibody (bottom parts of A, B and C). The PARP proenzyme (116 kD) is indicated in the top parts, and the cleaved subunit (85 kD) is indicated in the bottom parts (D). The levels of Cisplatin-induced MMP9 phosphorylation are indicated in the top parts of E, F and G, and the relative levels to the β-actin expression are indicated in the bottom parts of E, F and G. MMP-9 activity in Caov-3 cells was measured with a gelatin-zymography. The Pro-MMP9 is indicated in the top parts, and the Active-MMP9 is indicated in the bottom parts (H). The respective band densitometry analyses were performed using the Image J software program. The values represent the mean ± SE of at least three independent experiments. Significant differences are indicated by asterisks.**p < 0.01, * p < 0.05.