Figure 5

Cell-associated procoagulant activity and invasion in MDA-MB-231 cells, flTF expression and asTF mRNA transcription in SKOV-3 and OVCAR-3 cells. Panel a: Cell procoagulant activity. MDA-MB-231 cells were treated with 10 μM of PD98059, 10 μM of LY294002, 0.1 μM of wortmannin for 24 hrs and washed. Then clotting time was measured with these cells after the addition of human blood plasma and calcium. The results represent 3 different experiments. * : p<0.05 between PD98059 group versus LY294002 and wortmannin groups. Panel b: Cell invassive capacity : the same cells as in the panel a were cultured in the invasion chambers for 22 hrs. The cells penetrated through the matrigel were counted in microscopy. The data were from 3 independent experiments. * : P<0.05 in comparison to the control cells. Panel c: Regulation of TF protein in SKOV-3 and OVCAR-3 cells. SKOV-3 and OVCAR-3 cells were treated 10 μM of PD98059 with or without A6730 and EGFR siRNA for 24 hrs. The protein was extracted from the washed cells and examined for TF expression by western blot. The data were from 3 repeated blot experiments. Significant difference was found between the PD98059 group versus all of the other groups within the same cells (p<0.05). Panel d: asTF levels in PD98059 treated cells. The cells were treated by 10 μM of PD98059 with or without 0.1 μM erlotinib 10 μM A6730 and EGFR siRNA for 24 hrs. qPCR data were from 3 dependent experiments. * : p<0.05 for PD98059 groups in comparison with all of the other groups in the same cell lines.