Bombesin-Dependent Pro-MMP-9 Activation in Prostatic Cancer Cells Requires β1 Integrin Engagement

doi:10.1006/excr.2002.5609

Experimental Cell Research

Volume 280, Issue 1, 15 October 2002, Pages 1-11

https://doi.org/10.1006/excr.2002.5609 Get rights and content

Abstract

Bombesin-like peptides, including the mammalian homologue gastrin-releasing peptide, are highly expressed and secreted by neuroendocrine cells in prostate carcinoma tissues and are likely to be related to the progression of this neoplastic disease. Previously, we demonstrated that bombesin increased migration and protease expression in androgen-independent cells. In this work we show that bombesin is able to activate pro-MMP-9 through a mechanism involving the β1 integrin subunit. In fact, MMP-9 processing was evident only when β1 integrin was engaged with specific adhesive substrates, such as type I collagen, or when cells were seeded on dishes coated with antibodies against β1 integrin, resulting in activation of the surface ligand. When exogenous pro-MMP-9 was added to PC3 cells, MMP-9 active forms were produced within 30 min by bombesin-treated cultures while control cultures expressed activated forms only after a longer time and at lower levels. MMP-9 activation required cytoskeleton integrity since this effect was abolished by cytochalasin D. Engagement of β1 integrin caused an increased membrane-linked uPA activity which was required for MMP-9 activation. The cross talk between bombesin- and β1-integrin-engaged signals seems to be crucial for the modulation of both membrane-linked uPA activity and MMP-9 activation and triggers complex intracellular signaling pathways requiring activation of tyrosine kinase activity, including that of src and PI3K. The β1 integrin may be considered an important mechanism by which bombesin induces MMP-9 activation. This finding supports the idea that cellular responses to growth factors may be driven by cell–matrix interactions and stresses the role of neuroendocrine factors in prostate carcinoma progression.

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Further, we also showed that integrin β1 expression is particularly critical to GRP-R–mediated cell migration as evidenced by the significant decrease observed in neuroblastoma cell migration upon inhibition of integrin β1 with siRNA. It has been reported that GRP can stimulate cell migration in prostate13 and breast14 cancers through binding to its cognate receptor, GRP-R. Similar studies using the GRP analog, bombesin, have shown that bombesin/GRP-R signaling enhances the migratory and invasive capacity of colorectal carcinoma cells in a dose-dependent fashion,15 although the exact molecular mechanisms still remain unclear. While the role of GRP-R signaling in neuroblastoma tumorigenesis has been established,6 the effects of such signaling events as they relate to cell migration and invasion have not been examined.
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GRP increased neuroblastoma cell migration and expressions of MMP-2 whereas the TIMP-1 level decreased. GRP-R overexpression stimulated SK-N-SH cell migration and upregulated integrin α2, α3, and β1 protein as well as mRNA expression. Targeted silencing of integrin β1 inhibited cell migration.
GRP/GRP-R signaling contributes to neuroblastoma cell migration and invasion. Moreover, the integrin ß1 subunit critically regulates GRP-R–mediated neuroblastoma cell migration and invasion.
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The epidermal growth factor receptor (EGFR) family of receptor tyrosine kinases function as a common signaling conduit for membrane receptors that lack intrinsic enzymatic activity, such as G-protein coupled receptors and integrins. GPR30, an orphan member of the seven transmembrane receptor (7TMR) superfamily has been linked to specific estrogen binding, rapid estrogen-mediated activation of adenylyl cyclase and the release of membrane-tethered proHB-EGF. More recently, GPR30 expression in primary breast adenocarcinoma has been associated with pathological parameters commonly used to assess breast cancer progression, including the development of extramammary metastases. This newly appreciated mechanism of cross communication between estrogen and EGF is consistent with the observation that 7TMR-mediated transactivation of the EGFR is a recurrent signaling paradigm and may explain prior data reporting the EGF-like effects of estrogen. The molecular details surrounding GPR30-mediated release of proHB-EGF, the involvement of integrin β1 as a signaling intermediary in estrogen-dependent EGFR action, and the possible implications of these data for breast cancer progression are discussed herein.
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Matrix metalloproteinases (MMPs) are endopeptidases known to mediate acute neuronal injury, but it is unclear whether these proteases are induced by the primary insult or by inflammation associated with injury. We have reported recently that interleukin-1 (IL-1) induces neurotoxicity by an astrocyte-dependent mechanism. The aim of the present study was to test the hypothesis that MMPs mediate IL-1 neurotoxicity in rat, glial–neuronal cocultures. IL-1β induced the release of astrocytic MMP-9 in cocultures, whilst an antagonist of MMP-9 inhibited IL-1β-induced neuronal death. Urokinase plasminogen activator (uPA) was constitutively expressed on neuronal membrane fractions, and amiloride (an antagonist of uPA) or plasminogen activator inhibitor (PAI)-1 significantly reduced IL-1β-induced neurotoxicity. Thus, neuronal uPA contributes to IL-1 neurotoxicity, and may be responsible for activating MMP-9 released from IL-1-primed astrocytes. In summary, IL-1-induced neurotoxicity is dependent on extracellular protease activity, and these mechanisms may contribute to neuronal cell death in CNS diseases.
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BACKGROUND: Carcinoma of the prostate (CaP) is the most commonly diagnosed cancer in men in the United States. Signal transduction molecules such as tyrosine kinases play important roles in CaP. Src, a nonreceptor tyrosine kinase (NRTK) and the first proto-oncogene discovered is shown to participate in processes such as cell proliferation and migration in CaP. Underscoring NRTK's and, specifically, Src's importance in cancer is the recent approval by the US Food and Drug Administration of dasatinib, the first commercial Src inhibitor for clinical use in chronic myelogenous leukemia (CML). In this review we will focus on NRTKs and their roles in the biology of CaP. MATERIALS AND METHODS: Publicly available literature from PubMed regarding the topic of members of NRTKs in CaP was searched and reviewed. RESULTS: Src, FAK, JaK1/2, and ETK are involved in processes indispensable to the biology of CaP: cell growth, migration, invasion, angiogenesis, and apoptosis. CONCLUSIONS: Src emerges as a common signaling and regulatory molecule in multiple biological processes in CaP. Src's relative importance in particular stages of CaP, however, required further definition. Continued investigation of NRTKs will increase our understanding of their biological function and potential role as new therapeutic targets.
Bone microenvironment modulates expression and activity of cathepsin B in prostate cancer
2005, Neoplasia
Prostate cancers metastasize to bone leading to osteolysis. Here we assessed proteolysis of DOcollagen I (a bone matrix protein) and, for comparison, DO-collagen IV, by living human prostate carcinoma cells in vitro. Both collagens were degraded, this degradation was reduced by inhibitors of matrix metallo, serine, cysteine proteases. Because secretion of the cysteine protease cathepsin B is increased in human breast fibroblasts grown on collagen I gels, we analyzed cathepsin B levels and secretion in prostate cells grown on collagen I gels. Levels and secretion were increased only in DU145 cells-cells that expressed the highest baseline levels of cathepsin B. Secretion of cathepsin B was also elevated in DU145 cells grown in vitro on human bone fragments. We further investigated the effect of the bone microenvironment on cathepsin B expression and activity in vivo in a SCID-human model of prostate bone metastasis. High levels of cathepsin B protein and activity were found in DU145, PC3, LNCaP bone tumors, although the PC3 and LNCaP cells had exhibited low cathepsin B expression in vitro. Our results suggest that tumor-stromal interactions in the context of the bone microenvironment can modulate the expression of the cysteine protease cathepsin B.
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Citation Excerpt :
In our studies, the CRV peptide only weakly inhibited pro-MMP-2 activation, and the C domains of MMP-2 and MMP-9 did not compete with each other in binding assays. The finding that CRV mimics an integrin activation epitope provides an explanation for the requirement of ligand-engaged integrins in pro-MMP-9 activation (37, 40). We also demonstrate that uPAR, which is required for MMP-9 activation, associates with pro-MMP-9 in HT1080 and THP-1 cells.
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Regular ArticleBombesin-Dependent Pro-MMP-9 Activation in Prostatic Cancer Cells Requires β1 Integrin Engagement

Abstract

Eur. J. Cancer

Am. J. Pathol.

J. Biol. Chem.

J. Biol. Chem.

Curr. Opin. Cell Biol.

Curr. Opin. Cell Biol.

J. Biol. Chem.

Exp. Cell Res.

Cell

Blood

Exp. Cell. Res.

J. Biol. Chem.

Blood

J. Biol. Chem.

J. Biol. Chem.

J. Biol. Chem.

Neuroendocrine differentiation in human prostatic carcinoma

Hum. Pathol.

Acquisition of neuroendocrine characteristics by prostate tumor cells is reversible: Implications for prostate cancer progression

Cancer Res.

Autocrine and paracrine signaling through neuropeptide receptors in human cancer

Oncogene

Neuropeptide-induced androgen independent prostate cancer cells: Roles of nonreceptor tyrosine kinases etk/bmx, SRC, and focal adhesion kinase

Mol. Cell. Biol.

Bombesin stimulates growth of human prostatic cancer cells in vitro

Cancer

In vitro regulation of pericellular proteolysis in prostatic tumor cells treated with bombesin

Int. J. Cancer

Pro-gastrin-releasing peptide (ProGRP)—A useful marker in small cell lung carcinomas

Anticancer Res.

Regular Article
Bombesin-Dependent Pro-MMP-9 Activation in Prostatic Cancer Cells Requires β1 Integrin Engagement