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M513025200v1
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Papers In Press, published online ahead of print May 4, 2006
J. Biol. Chem, 10.1074/jbc.M513025200
Submitted on December 6, 2005
Revised on April 27, 2006
Accepted on May 4, 2006

Annexin I regulates SKCO-15 cell invasion by signaling through formyl peptide receptors

Brian A. Babbin, Winston Y. Lee, Charles A. Parkos, L.Matthew Winfree, Adil Akyildiz, Mauro Perretti, and Asma Nusrat

Pathology & Laboratory Medicine, Emory University, Atlanta, GA 30322

Corresponding Author: bbabbin{at}emory.edu

Annexin 1 (AnxA1) is a multifunctional phospholipid binding protein associated with the development of metastasis in some invasive epithelial malignancies. However, the role of AnxA1 in the migration/invasion of epithelial cells is not known. In this study, experiments were performed to investigate the role of AnxA1 in the invasion of a model epithelial cell line, SKCO-15, derived from colorectal adenocarcinoma. siRNA-mediated knockdown of AnxA1 expression resulted in a significant reduction in invasion through matrigel-coated filters. Localization studies revealed a translocation of AnxA1 to the cell surface upon the induction of cell migration, and functional inhibition of cell surface AnxA1 using antiserum (LCO1) significantly reduced cell invasion. Conversely, SKCO-15 cell invasion was increased by approximately two fold in the presence of recombinant full length AnxA1 and the AnxA1 N-terminal derived peptide mimetic, Ac2-26. Since extracellular AnxA1 has been shown to regulate leukocyte migratory events through interactions with n-formyl peptide receptors (nFPRs), we examined the expression of FPR-1, FPRL-1, and FPRL-2 in SKCO-15 cells by RT-PCR and identified expression of all three receptors in this cell line. Treatment of SKCO-15 cells with AnxA1, Ac2-26, and the classical nFPR agonist, fMLP, induced intracellular calcium release consistent with nFPR activation. Furthermore, the nFPR antagonist, Boc2, abrogated the AnxA1 and Ac2-26- induced intracellular calcium release and increase in SKCO-15 cell invasion. Together, these results support an autocrine/paracrine role for membrane AnxA1 in stimulating SKCO-15 cell migration through nFPR activation. The findings in this study suggest that activation of nFPRs stimulates epithelial cell motility important in the development of metastasis as well as wound healing.


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