Characterization of Functional Thrombin Receptors in Human Pancreatic Tumor Cells (MIA PACA-2)

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Abstract

Summary

In this article, the “tethered ligand” thrombin receptor was identified on human pancreatic tumor cells, MIA PaCa-2, using immunofluorescence studies with a monoclonal anti-thrombin receptor antibody. Pharmacological characterization, using 3H-labeled thrombin receptor activating peptide-6 (TRAP-6) as radioligand, demonstrated a single class of high-affinity binding sites (KD = 9.1 ± 1.8 × 10−7 M) and a binding capacity of 13.9 ± 0.7 fmol/mg protein. These binding sites represent functional thrombin receptors, as shown by α-thrombin- and TRAP-6-induced mobilization of free intracellular calcium, protein kinase C translocation from cytosol to the cell membrane, and stimulation of DNA synthesis in MIA PaCa-2 cells. These results provide the first identification of tethered ligand thrombin receptor in human pancreatic cancer cells and suggest thrombin receptor involvement in mechanisms of human pancreatic tumor progression.

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