Cysteine Cathepsins Are Central Contributors of Invasion by Cultured Adenosylmethionine Decarboxylase-Transformed Rodent Fibroblasts

Ravanko, Kirsi; Järvinen, Kristiina; Helin, Jari; Kalkkinen, Nisse; Hölttä, Erkki

doi:10.1158/0008-5472.can-03-2993

Cited by 45 publications

(45 citation statements)

References 47 publications

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“…Conversely, cosilencing either uPAR or ␣V integrin in M6P/IGF2R-silenced cells revealed that the enhanced adhesion to vitronectin was mediated by ␣V integrin and not uPAR. A direct and strong function of uPAR in tumor cell adhesion to vitronectin is therefore not supported by our results, which is also in agreement with the findings of a recent report (Smith et al, 2008).Various cathepsins promote tumor invasiveness directly via degradation of extracellular matrix components or via activation of other proteinases, such as pro-uPA (Kobayashi et al, 1991;Ravanko et al, 2004). In addition, autocrine stimulation by IGF2 or TGF␤ might influence the Plg activation system (Leivonen and Kahari, 2007;Samani et al, 2007).…”

supporting

confidence: 86%

Mannose 6-Phosphate/Insulin-like Growth Factor 2 Receptor Limits Cell Invasion by Controlling αVβ3 Integrin Expression and Proteolytic Processing of Urokinase-type Plasminogen Activator Receptor

Schiller

Szekeres

Binder

et al. 2009

MBoC

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The multifunctional mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R) is considered a tumor suppressor. We report here that RNA interference with M6P/IGF2R expression in urokinase-type plasminogen activator (uPA)/urokinase-type plasminogen activator receptor (uPAR) expressing human cancer and endothelial cells resulted in increased pericellular plasminogen activation, cell adhesion, and higher invasive potential through matrigel. M6P/IGF2R silencing led also to the cell surface accumulation of urokinase and plasminogen and enhanced expression of ␣V integrins. Genetic rescue experiments and inhibitor studies revealed that the enhanced plasminogen activation was due to a direct effect of M6P/IGF2R on uPAR, whereas increased cell adhesion to vitronectin was dependent on ␣V integrin expression and not uPAR. Increased cell invasion of M6P/IGF2R knockdown cells was rescued by cosilencing both uPAR and ␣V integrin. Furthermore, we found that M6P/IGF2R expression accelerates the cleavage of uPAR. M6P/IGF2R silencing resulted in an increased ratio of full-length uPAR to the truncated D2D3 fragment, incapable of binding most uPAR ligands. We conclude that M6P/IGF2R controls cell invasion by regulating ␣V integrin expression and by accelerating uPAR cleavage, leading to the loss of the urokinase/vitronectin/integrin-binding site on uPAR. INTRODUCTIONThe loss of the mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF2R, CD222) has been described in many human malignancies, and it is considered a tumor suppressor (Scott and Firth, 2004;Hebert, 2006). M6P/IGF2R is a multifunctional receptor possessing distinct binding sites for structurally unrelated ligands and membrane partners (Ghosh et al., 2003). It is not clear, however, what functional consequence of the loss of M6P/IGF2R is directly and determinedly responsible for tumor progression in vivo. First, the M6P/IGF2R-mediated degradation of insulin-like growth factor (IGF) 2 may play a role in suppression of tumor growth (Samani et al., 2007). Second, the lack of M6P/IGF2R in tumor cells may also contribute to their higher invasiveness due to impaired intracellular trafficking of cathepsins (Lorenzo et al., 2000), observed also in M6P/ IGF2R-deficient cell lines (Kasper et al., 1996). Third, some of the M6P/IGF2R's ligands, such as retinoic acid and granzyme B, were shown to induce apoptosis via the receptor (Kang et al., 1997;Motyka et al., 2000). Fourth, the loss of M6P/IGF2R may reduce the availability of active transforming growth factor (TGF)-␤ and thus its inhibitory effects on cell proliferation (Dennis and Rifkin, 1991;Godar et al., 1999;Leksa et al., 2005). Finally, M6P/IGF2R binds the urokinasetype plasminogen activator receptor (uPAR, CD87) and plasminogen (Plg), two components of the fibrinolytic system (Nykjaer et al., 1998;Godar et al., 1999;Leksa et al., 2002;Kreiling et al., 2003). Plg activation is crucial to direct cell migration through tissue barriers. On binding to uPAR, pro-urokinase (pro-uPA) is proteolytically ac...

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supporting

confidence: 86%

Mannose 6-Phosphate/Insulin-like Growth Factor 2 Receptor Limits Cell Invasion by Controlling αVβ3 Integrin Expression and Proteolytic Processing of Urokinase-type Plasminogen Activator Receptor

Schiller

Szekeres

Binder

et al. 2009

MBoC

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“…We have previously shown that cysteine cathepsin L, a lysosomal protease, is the major secreted protease responsible for the invasive activity of Amdc cells. 26 Notably, cathepsin L, which is capable of degrading both basement membrane and extracellular matrix (ECM) proteins, 29 has also been identified as a target of v-Fos/AP-1. 20 In addition, we have found thymosin b4, a major actin sequestering protein, to be important for the regulation of the morphology and migration of Amdc cells.…”

Section: Discussionmentioning

confidence: 99%

“…26 As we here found also integrin a6 to be necessary for the invasiveness of these cells, we studied whether the c-Jun dependent regulation of the activity of cathepsin L might be mediated by integrin a6. Thus, integrin a6 function was blocked with the neutralizing antibody in cell culture and the cellular RNAs and secreted proteins were collected for the analysis of cathepsin L mRNA and protein/activity levels.…”

Section: C-jun-induced Upregulation Of Cathepsin L Is Independent Of mentioning

confidence: 94%

“…21,26 After trypsinization and before the invasion assays, 15,000 Amdc cells or 30,000 HT-1080 cells were preincubated with 10 lg/ml anti-integrin a6, b1 or b7, or isotype control antibody for 30 min. The same concentration of antibodies was also used in the Matrigel layers.…”

Section: Matrigel Invasion Assaymentioning

confidence: 99%

“…To get free of the added antibodies, the media were filtered using Amicon 100 kDa Ultra filter devices (Millipore, Billerica, MA), and then the proteins were concentrated and the growth medium changed to acidic activation buffer (0.1 M sodium acetate pH 5.0, 1 mM EDTA, 0.01% Triton X-100) using 5 kDa Ultra filter devices. Finally, the activity of secreted cathepsin L was assayed as previously reported, 26 in the presence of heparin.…”

Section: Assay Of Cathepsin L Activitymentioning

confidence: 99%

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Identification of integrins α6 and β7 as c‐Jun‐ and transformation‐relevant genes in highly invasive fibrosarcoma cells

Kielosto

Nummela

Järvinen

et al. 2009

Intl Journal of Cancer

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Understanding the mechanisms of tumor cell invasion is essential for our attempts to prevent cancer deaths. We screened by DNA microarrays the c-Jun-and transformation-related gene expression changes in S-adenosylmethionine decarboxylase (AdoMetDC)-overexpressing mouse fibroblasts that are highly invasive in vivo, and their derivatives expressing a tetracycline-inducible dominant-negative mutant of c-Jun (TAM67) or c-Jun shRNA. Among the small set of target genes detected were integrins a6 and b7, cathepsin L and thymosin b4, all upregulated in the AdoMetDC-transformed cells and downregulated upon reversal of transformation by TAM67 or c-Jun shRNA. The upregulation of integrin a6 subunit, pairing with integrin b1, endowed the transformed cells with the capability to attach to basement membrane laminin and to spread. Further, inhibition of integrin a6 or b1 function with neutralizing antibodies blocked the invasiveness of AdoMetDC-transformants and human HT-1080 fibrosarcoma cells in three-dimensional Matrigel. Moreover, immunohistochemical analyses showed strong integrin a6 staining in high-grade human fibrosarcomas. Our data show that c-Jun can regulate all three key steps of invasion: cell adhesion (integrin a6), basement membrane/extracellular matrix degradation (cathepsin L) and cell migration (thymosin b4). In addition, this is the first study to associate integrin b7, known as a leukocyte-specific integrin binding to endothelial/epithelial cell adhesion molecules, with the transformed phenotype in cells of nonleukocyte origin. As tumor cell invasion is a prerequisite for metastasis, the observed critical role of integrin a6b1 in fibrosarcoma cell invasion/spreading allures testing antagonists to integrin a6b1, alone or combined with inhibitors of cathepsin L and thymosin b4, as chemotherapeutic agents. ' 2009 UICC

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Prolyl 4‐hydroxylase subunit alpha 1 (P4HA1) is a biomarker of poor prognosis in primary melanomas, and its depletion inhibits melanoma cell invasion and disrupts tumor blood vessel walls

et al. 2020

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Melanoma is an unpredictable, highly metastatic malignancy, and treatment of advanced melanoma remains challenging. Novel molecular markers based on the alterations in gene expression and the molecular pathways activated or deactivated during melanoma progression are needed for predicting the course of the disease already in primary tumors and for providing new targets for therapy. Here, we sought to identify genes whose expression in primary melanomas correlate with patient disease‐specific survival using global gene expression profiling. Many of the identified potential markers of poor prognosis were associated with the epithelial–mesenchymal transition, extracellular matrix formation, and angiogenesis. We studied further the significance of one of the genes, prolyl 4‐hydroxylase subunit alpha 1 (P4HA1), in melanoma progression. P4HA1 depletion in melanoma cells reduced cell adhesion, invasion, and viability in vitro. In melanoma xenograft assays, we found that P4HA1 knockdown reduced melanoma tumor invasion as well as the deposition of collagens, particularly type IV collagen, in the interstitial extracellular matrix and in the basement membranes of tumor blood vessels, leading to vessel wall rupture and hemorrhages. Further, P4HA1 knockdown reduced the secretion of collagen triple helix repeat containing 1 (CTHRC1), an important mediator of melanoma cell migration and invasion, in vitro and its deposition around tumor blood vessels in vivo. Taken together, P4HA1 is an interesting potential prognostic marker and therapeutic target in primary melanomas, influencing many aspects of melanoma tumor progression.

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Cysteine Cathepsins Are Central Contributors of Invasion by Cultured Adenosylmethionine Decarboxylase-Transformed Rodent Fibroblasts

Cited by 45 publications

References 47 publications

Mannose 6-Phosphate/Insulin-like Growth Factor 2 Receptor Limits Cell Invasion by Controlling αVβ3 Integrin Expression and Proteolytic Processing of Urokinase-type Plasminogen Activator Receptor

Mannose 6-Phosphate/Insulin-like Growth Factor 2 Receptor Limits Cell Invasion by Controlling αVβ3 Integrin Expression and Proteolytic Processing of Urokinase-type Plasminogen Activator Receptor

Identification of integrins α6 and β7 as c‐Jun‐ and transformation‐relevant genes in highly invasive fibrosarcoma cells

Prolyl 4‐hydroxylase subunit alpha 1 (P4HA1) is a biomarker of poor prognosis in primary melanomas, and its depletion inhibits melanoma cell invasion and disrupts tumor blood vessel walls

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