Neutral endopeptidase inhibits prostate cancer cell migration by blocking focal adhesion kinase signaling

Makoto Sumitomo, Ruoqian Shen, Marc Walburg, Jie Dai, Yiping Geng, Daniel Navarro, Guy Boileau, Christos N. Papandreou, Filippo G. Giancotti, Beatrice Knudsen, David M. Nanus

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Abstract

Neutral endopeptidase 24.11 (NEP, CD10) is a cell-surface enzyme expressed by prostatic epithelial cells that cleaves and inactivates neuropeptides implicated in the growth of androgen-independent prostate cancer (PC). NEP substrates such as bombesin and endothelin-1 induce cell migration. We investigated the mechanisms of NEP regulation of cell migration in PC cells, including regulation of phosphorylation on tyrosine of focal adhesion kinase (FAK). Western analyses and cell migration assays revealed an inverse correlation between NEP expression and the levels of FAK phosphorylation and cell migration in PC cell lines. Constitutively expressed NEP, recombinant NEP, and induced NEP expression using a tetracycline-repressive expression system inhibited bombesin- and endothelin-1-stimulated FAK phosphorylation and cell migration. This results from NEP-induced inhibition of neuropeptide-stimulated association of FAK with cSrc protein. Expression of a mutated catalytically inactive NEP protein also resulted in partial inhibition of FAK phosphorylation and cell migration. Coimmunoprecipitation experiments show that NEP associates with tyrosine-phosphorylated Lyn kinase, which then binds the p85 subunit of phosphatidylinositol 3-kinase (PI3-K) resulting in an NEP-Lyn-PI3-K protein complex. This complex competitively blocks FAK-PI3-K interaction, suggesting that NEP protein inhibits cell migration via a protein-protein interaction independent of its catalytic function. These experiments demonstrate that NEP can inhibit FAK phosphorylation on tyrosine and PC cell migration through multiple pathways and suggest that cell migration which contributes to invasion and metastases in PC cells can be regulated by NEP.

Original languageEnglish
Pages (from-to)1399-1407
Number of pages9
JournalJournal of Clinical Investigation
Volume106
Issue number11
DOIs
Publication statusPublished - 2000

All Science Journal Classification (ASJC) codes

  • Medicine(all)

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    Sumitomo, M., Shen, R., Walburg, M., Dai, J., Geng, Y., Navarro, D., Boileau, G., Papandreou, C. N., Giancotti, F. G., Knudsen, B., & Nanus, D. M. (2000). Neutral endopeptidase inhibits prostate cancer cell migration by blocking focal adhesion kinase signaling. Journal of Clinical Investigation, 106(11), 1399-1407. https://doi.org/10.1172/JCI10536