Figure 9.

EGF signaling contributes to KLK8-induced activation of PI3K-Akt-mTOR signaling pathway and KLK8-induced pro-proliferation and anti-apoptotic effects in pancreatic cancer cells. Mia-paca-2 and Panc-1 cells were treated with EGFR antagonist AG1478 (100 nM). Twenty-four hours later, cells were harvested for measuring cell protein level, proliferation and apoptosis. (A) Expression level of EGF in Mia-paca-2 and Panc-1 cells were evaluated by ELISA assay. (B) Levels of key proteins in PI3K-AKT-mTOR signaling pathways were examined in Mia-paca-2 and Panc-1 cells using western blot (C) Cell proliferation was detected by CCK8 assay in Mia-paca-2 and Panc-1 cells. (D) Colony formation was detected in Mia-paca-2 and Panc-1 cells. (E) Demonstrated the quantified data of cell colonies. (F) Cell apoptosis was determined by Annexin V-FITC and PI double staining analysis performed by flow cytometry. Representative flow cytometry images were shown. (G) Demonstrated the quantified data of cell apoptosis. (H) Pro-apoptotic markers cleaved caspase-3, cleaved caspase-9 and Bax assessed by western blot. Data were presented as the mean ± SEM (n=3). **p <0.01 vs Lv-control; p < 0.05, ^##p < 0.01 vs Lv-KLK8.