Effect of LY294002 on HPV-16 E7-induced HIF-1α, VEGF, and IL-8 expression and <i>in vitro</i> angiogenesis in NSCLC cells.

<p>HPV-16 E7-transfected NSCLC cells were pretreated for 24 h with different concentrations of LY294002. (A) HIF-1α and p-Akt protein levels in transfected NSCLC cells (Left: A549, Right: NCI-H460) were analyzed by Western blotting. (B) VEGF and IL-8 protein concentration in the conditioned media derived from transfected A549 cells was determined by ELISA. (C) VEGF and IL-8 mRNA levels in transfected A549 cells were determined by real-time PCR. (D) Results <i>in </i><i>vitro</i> angiogenesis (A549 cells). HUVECs (5×10³cells/well) were seeded onto the surface of 96-well cell culture plates pre-coated with polymerized ECMatrix and then incubated at 37°C for 6 to 8 h in the conditioned media derived from HPV-16 E7-transfected A549 cells in the absence or presence of LY294002. Left: The tube formation was observed under a phase-contrast microscope (20×). Right: The total tube length in 3 random view-fields per well was by Scion image software measured and average value was calculated. All data are expressed as mean ± SD of three independent experiments. *<i>P</i><0.05, **<i>P</i><0.01.