Figure 3

HCCR-1 overexpression is mediated with EGF-induced PI3K/AKT/mTOR signaling. Western blot analysis of HCCR-1 expression in PANC-1 cells stimulated with EGF. A) Dose dependency (at 24 h). Total protein were extracted from serum starved PANC-1 cells treated with EGF at different concentration, and subjected to Western blotting. B) Time course (at 100 ng/ml). Total protein were isolated from PANC-1 cells for Western blotting after different duration of EGF. EGF stimulates HCCR-1 expression in PANC-1 cells in a dose- and time-dependent manner. Tubulin was used as a loading control. Blocking PI3K or mTOR signaling with either C) LY294002 (50 μg/mL) or D) rapamycin (10 ng/ml), respectively, suppresses the EGF (100 ng/mL)-mediated HCCR-1 expression in PANC-1 cells. E) EGF induces the activity of Akt within 2 min at highest level with gradual decrease.