Ligand activation of alternatively spliced fibroblast growth factor receptor-1 modulates pancreatic adenocarcinoma cell malignancy

Pancreatic adenocarcinoma continues to be a devastating tumor (28,000 new cases per year in the United States; 10% 2-year survival). Pancreatic adenocarcinoma frequently (90% of the time) overexpresses fibroblast growth factor ligands (FGF-1 and FGF-2) and alternatively spliced high-affinity receptors (FGFR-ls) (FGFR-lct was previously found in normal pancreatic tissue). To study the significance of this observation in vitro, PANC-1 cells were stably transfected via the pMEXneo vector containing FGFR-la (PANC-la) or FGFR-ls (PANC-ls) isoforms. Cells were treated with 1 mg/ml of 5-fluorouracil. Cells were evaluated for growth inhibition, apoptosis (propidium iodide staining and flow cytometry, caspase 3 activation) and for Bcl-xL/BAX expression (by Western blot analysis). In vivo, 7 X 106 cells of each isoform were injected into nude Balb/c mice for xenograft formation (N = 10). Compared to PANC-ls (9%) in vitro, 5-fluorouracil-induced death was significantly (P < 0.05) increased in PANC-la (20%) at 24 hours. Increased cell death in PANC-la was mediated by activated caspase 3 and was correlated with decreased expression of Bcl-xL/BAX. In vivo, PANC-ls readily demonstrated formation of tumor xenograft at 2 weeks, whereas PANC-la did not form tumors. Alternative splicing of FGFR-1 to the s isoform appears to correlate with pancreatic adenocarcinoma cell growth in vivo and resistance to chemotherapy. Inhibition of FGFR-1 splicing or overexpression of FGFR-la inhibits pancreatic adenocarcinoma cell growth in vivo and restores cytotoxic responses to chemotherapy, thereby suggesting the basis of rational interventional strategies for this devastating tumor.