Tumor cell invasion of model 3-dimensional matrices: demonstration of migratory pathways, collagen disruption, and intercellular cooperation

Horino, Kei; Kindezelskii, Andrei L.; Elner, Victor M.; Hughes, Bret A.; Petty, Howard R.

doi:10.1096/fj.00-0392com

Cited by 27 publications

(18 citation statements)

References 17 publications

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“…It has been demonstrated that uPAR-positive cells express higher invasive capacity than uPAR-negative cells on gelatin-coated filters. 41 However, the stable expression of uPAR in the presence of SCH-66336 suggests that other factors are involved in the reduced migration of PC-3 cells on filters coated with gelatin. The lower activities of uPA, plasmin and gelatinases in cancer cells treated with SCH-66336 could contribute to reduced migratory capacity.…”

Section: Discussionmentioning

confidence: 99%

Farnesyltransferase inhibitor SCH‐66336 downregulates secretion of matrix proteinases and inhibits carcinoma cell migration

Desrosiers

Cusson

Turcotte

et al. 2005

Intl Journal of Cancer

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The ras oncogenes are among those most frequently found in human cancers. Blocking Ras farnesylation is a promising strategy for arresting cancer growth. Ras activates several signaling pathways with key roles in cellular proliferation, invasion, metastasis and angiogenesis. Furthermore, proteolytic activities of matrix proteinases such as urokinase-type plasminogen activator (uPA) and matrix metalloproteinases (MMPs) are regulated by Ras isoforms. Thus, we investigated the effects of SCH-66336, a farnesyltransferase inhibitor, on secretion of components of the plasminogen activation system as well as on the gelatinases MMP-2 and MMP-9, which play pivotal roles in matrix remodeling. SCH-66336 up to 5 M did not significantly alter the viability of prostate (PC-3) and renal (Caki-1) cancer cells incubated in serum-depleted medium. SCH-66336 partly inhibited the processing of H-Ras, while levels of mature N-Ras and K-Ras remained unaffected. Under these noncytotoxic conditions, uPA and tPA levels were lowered in culture medium but raised in cell lysates, suggesting inhibition of trafficking pathways. In contrast, SCH-66336 had no effect on uPAR expression or on secreted PAI-1 levels. As expected, the reduction of uPA and tPA activities by SCH-66336 inhibited the conversion of plasminogen to plasmin by about 25% in PC-3 cells. SCH-66336 also inhibited the levels of secreted pro-MMP-2 and pro-MMP-9 as well as the release of their inhibitors TIMP-1 and TIMP-2. SCH-66336 decreased both the adhesion and even more so the migration of PC-3 cells on gelatin. Thus, SCH-66336 inhibited farnesylation in both cancer cell types, and H-Ras functions should be reduced by the drug. In addition, the lower levels of secreted proteinases in the presence of SCH-66336 suggest that reduced matrix remodeling and cell migration should occur in treated tumors.

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Section: Discussionmentioning

confidence: 99%

Farnesyltransferase inhibitor SCH‐66336 downregulates secretion of matrix proteinases and inhibits carcinoma cell migration

Desrosiers

Cusson

Turcotte

et al. 2005

Intl Journal of Cancer

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“…DQ-collagen type IV has been mainly applied to cells cultured on matrices containing the substrate (Horino et al 2001;Sameni et al 2001;Elner 2002;Frederiks, Mook Premzl et al 2003). Invasion of fibrosarcoma cells in gelatin matrix was accompanied by collagenolytic activity due to MMP activity (Horino et al 2001).…”

Section: In Situ Zymography With Other Quenched Fluorogenic Substratesmentioning

confidence: 99%

“…Invasion of fibrosarcoma cells in gelatin matrix was accompanied by collagenolytic activity due to MMP activity (Horino et al 2001). Sameni et al (2000Sameni et al ( ,2001 investigated the site of proteolysis in a three-dimensional gelatin matrix embedded with DQ-collagen type IV and human cancer cells cultured on top of it.…”

Section: In Situ Zymography With Other Quenched Fluorogenic Substratesmentioning

confidence: 99%

“…Figure 5 shows the localization of fluorescent breakdown products of Bodipy-casein in colon cancer metastases of mouse liver. DQ-BSA has also been used instead of DQ-casein as substrate for the localization of uPA activity in matrices containing fibrosarcoma cells (Horino et al 2001) and retinal pigment cells (Elner 2002). The specificity of extracellular proteolysis of BSA by uPA was assessed by inhibition with PAI-1.…”

Section: In Situ Zymography With Other Quenched Fluorogenic Substratesmentioning

confidence: 99%

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Metabolic Mapping of Proteinase Activity with Emphasis on In Situ Zymography of Gelatinases

Frederiks

Mook

2004

J Histochem Cytochem.

110

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S U M M A R Y Proteases are essential for protein catabolism, regulation of a wide range of biological processes, and in the pathogenesis of many diseases. Several techniques are available to localize activity of proteases in tissue sections or cell preparations. For localization of the activity of matrix metalloproteinases, in situ zymography was introduced some decades ago. The procedure is based on zymography using SDS polyacrylamide gels containing gelatin, casein, or fibrin as substrate. For in situ zymography, either a photographic emulsion containing gelatin or a fluorescence-labeled proteinaceous macromolecular substrate is brought into contact with a tissue section or cell preparation. After incubation, enzymatic activity is revealed as white spots in a dark background or as black spots in a fluorescent background. However, this approach does not allow precise localization of proteinase activity because of limited sensitivity. A major improvement in sensitivity was achieved with the introduction of dye-quenched (DQ-)gelatin, which is gelatin that is heavily labeled with FITC molecules so that its fluorescence is quenched. After cleavage of DQ-gelatin by gelatinolytic activity, fluorescent peptides are produced that are visible against a weakly fluorescent background. The incubation with DQ-gelatin can be combined with simultaneous immunohistochemical detection of a protein on the same section. To draw valid conclusions from the findings with in situ zymography, specific inhibitors need to be used and the technique has to be combined with immunohistochemistry and zymography. In that case, in situ zymography provides data that extend our understanding of the role of specific proteinases in various physiological and pathological conditions.

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“…21,22 Supporting this, in vitro experiments have shown that noninvasive cancer cells can progress through three-dimensional matrices following the track of invasive tumor cells, which suggests that tumor cell migration and invasion may be modulated by signals generated by other tumor cells. 23 Infiltrating central nervous system tumors give rise to signals, which can be read and followed by specific neural cell types, 24 underscoring the role of tumor-host signals for recruiting other cells into the metastases environment. It is reasonable to assume that transducing tumor cells with genes implicated in the metastatic process would likely enhance the targeting efficacy.…”

Section: Discussionmentioning

confidence: 99%

Tumor cells as cellular vehicles to deliver gene therapies to metastatic tumors

García‐Castro

Martínez-Palacio

Lillo³

et al. 2005

Cancer Gene Ther

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A long-pursued goal in cancer treatment is to deliver a therapy specifically to metastases. As a result of the disseminated nature of the metastatic disease, carrying the therapeutic agent to the sites of tumor growth represents a major step for success. We hypothesized that tumor cells injected intravenously (i.v.) into an animal with metastases would respond to many of the factors driving the metastatic process, and would target metastases. Using a model of spontaneous metastases, we report here that i.v. injected tumor cells localized on metastatic lesions. Based on this fact, we used genetically transduced tumor cells for tumor targeting of anticancer agents such as a suicide gene or an oncolytic virus, with evident antitumoral effect and negligible systemic toxicity. Therefore, autologous tumor cells may be used as cellular vehicles for systemic delivery of anticancer therapies to metastatic tumors. Cancer Gene Therapy ( Keywords: neoplasm metastasis; delivery system; oncolytic virus; NOD SCID mice C ancer is often diagnosed when the disease has already disseminated and most of the patient deaths are related to metastatic disease. Current treatment options for metastatic tumors lack efficacy and metastases targeting remains a major challenge for curing cancer. Novel cell-and gene-based therapies have been developed aimed to deliver an antitumor effect specifically to metastases. Some strategies rely on the immune system for selectivity, such as the stimulation of effector immune cells 1 or the use of monoclonal antibodies that recognize tumor antigens. 2 Others target angiogenic 3-5 or anaerobic signals 6,7 that arise at the metastases. Recently, oncotropic viruses have been used to selectively kill the tumor cells with the advantage that once targeting is achieved the propagation of the virus amplifies the therapy. 8,9 These promising new therapies have fallen behind expectations mainly because of their limited capacity for effectively targeting in vivo. 2,10 The process of metastasis is a turning point in the progression of malignant solid tumors. Million tumor cells leave the primary tumor, reach the bloodstream and disseminate through the body. 11 Only a minority will eventually survive to become the origin of a new cancer nodule 12 at anatomical sites that are specific for the tumor type. 13 The dissemination of cancer cells from the primary tumor and their homing in specific organs involve several steps: invasion, detachment, circulation, cell adhesion, motility and invasion again. Cells must express a specific receptor molecule repertoire (cell adhesion molecules, chemokine receptors or integrin ligands among others) to complete this metastatic process. [14][15][16][17] We hypothesized that cancer cells may be good candidates to target established metastases in vivo because they express the receptor and effector molecules involved in the metastatic process. Autologous intravenously (i.v.) injected tumor cells should respond to metastasis-related cues as the cells leaving the primary tumor did when th...

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Tumor cell invasion of model 3-dimensional matrices: demonstration of migratory pathways, collagen disruption, and intercellular cooperation

Cited by 27 publications

References 17 publications

Farnesyltransferase inhibitor SCH‐66336 downregulates secretion of matrix proteinases and inhibits carcinoma cell migration

Farnesyltransferase inhibitor SCH‐66336 downregulates secretion of matrix proteinases and inhibits carcinoma cell migration

Metabolic Mapping of Proteinase Activity with Emphasis on In Situ Zymography of Gelatinases

Tumor cells as cellular vehicles to deliver gene therapies to metastatic tumors

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