Detection of plasminogen activators in oral cancer by laser capture microdissection combined with zymography

Curino, Alejandro Carlos; Patel, Vyomesh; Nielsen, Boye Schnack; Iskander, Andrew; Ensley, John F.; Yoo, George H.; Holsinger, F. Christopher; Myers, Jeffrey N.; El-Nagaar, Adel; Kellman, Robert M.; Shillitoe, Edward J.; Molinolo, Alfredo A.; Gutkind, J. Silvio; Bugge, Thomas H.

doi:10.1016/j.oraloncology.2004.05.011

Cited by 20 publications

(15 citation statements)

References 23 publications

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“…OSCC cell lines display enhanced uPA expression relative to cells derived from normal oral mucosa and exhibit increased uPA-dependent invasion in vitro [10–12]. Explants of OSCC tissues also show higher levels of uPA relative to adjacent non-malignant tissues [13,14]. …”

Section: Introductionmentioning

confidence: 99%

Urinary-Type Plasminogen Activator Receptor/α3β1 Integrin Signaling, Altered Gene Expression, and Oral Tumor Progression

Ghosh

Koblinski

Johnson

et al. 2010

Molecular Cancer Research

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Oral squamous cell carcinoma (OSCC) has 50% 5-year survival rate, highlighting our limited understanding of the molecular events that contribute to disease progression. Microarray analyses of primary oral tumors have identified urinary-type plasminogen activator (uPA) and its receptor (uPAR) as key genes associated with human OSCC progression. The uPAR functions as both a proteinase receptor and an integrin ligand, modifying proteolysis, migration, integrin signaling, and cellular transcription. In the current study, uPAR expression levels were modified in OSCC cells followed by analysis of tumor growth in an in vivo orthotopic xenograft model and by transcriptional profiling. Overexpression of uPAR resulted in more infiltrative and less differentiated tumors, with ill-defined borders, cytologic atypia, and enhanced vascularity. Analysis of serial sections of both murine experimental tumors and microarrayed human OSCC showed a statistically significant association between uPAR and α 3 integrin colocalization in areas exhibiting extracellular signal-regulated kinase phosphorylation, suggesting that uPAR/α 3 integrin interaction potentiates extracellular signal-regulated kinase signaling in vivo. This is supported by cDNA microarray analysis, which showed differential expression of 148 genes (113 upregulated and 35 downregulated). Validation of gene expression changes in human OSCC using immunohistochemistry and quantitative real-time PCR showed increased growth factors, proteinases/inhibitors, and matrix components in uPAR-overexpressing tumors. Together, these results support a model wherein increased uPAR expression promotes α 3 β 1 integrin association, resulting in increased mitogen-activated protein kinase signaling and transcriptional activation, leading to the formation of more aggressive tongue tumors. This combined approach has efficacy to identify additional biomarkers and/or prognostic indicators associated with aggressive human OSCC. Mol Cancer Res; 8(2); 145-58. ©2010 AACR.

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Section: Introductionmentioning

confidence: 99%

Urinary-Type Plasminogen Activator Receptor/α3β1 Integrin Signaling, Altered Gene Expression, and Oral Tumor Progression

Ghosh

Koblinski

Johnson

et al. 2010

Molecular Cancer Research

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“…Furthermore, it has been shown that high tumor PAI-1 levels, similarly to high uPA levels, predict a poor prognosis (Andreasen et al, 2000;Durand et al, 2004). PAI-1 is expressed in oral squamous cell carcinomas but not in normal oral mucosa; furthermore, PAI-1 is mainly expressed in cancer cells in oral carcinoma (Clayman et al, 1993;Nozaki et al, 1998;Curino et al, 2004;Lindberg et al, 2006). In this respect, oral carcinomas are different from other cancer types, such as breast, prostate, and colon cancers, in which PAI-1 is predominantly found in stromal myofibroblasts and not in the cancer cells themselves (Offersen et al, 2003;Illemann et al, 2004;Usher et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Epigenetic alterations of the SERPINE1 gene in oral squamous cell carcinomas and normal oral mucosa

Gao

Nielsen

Krogdahl

et al. 2010

Genes Chromosomes & Cancer

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A high level of plasminogen activator inhibitor-1 (PAI-1 or SERPINE1) in tumor extracts is a marker of a poor prognosis in human cancers, including oral carcinomas. However, the mechanisms responsible for the upregulation of PAI-1 in cancers remain unclear. Investigating specific PAI-1 expressing cells in oral carcinomas by immunohistochemistry, we found that PAI-1 was expressed in 18 of the 20 patients, mainly by cancer cells. Two showed PAI-1 positive stromal cells surrounding the tumor areas and five showed PAI-1 positive cells in tumor-adjacent normal epithelium. By real-time RT-PCR analysis, 17 of 20 patients with oral carcinoma were found to have between 2.5- and 50-fold increased tumor PAI-1 mRNA level, as compared with the matched tumor-adjacent normal tissues. The PAI-1 mRNA level in connective tissues from 15 healthy volunteers was similar to the level in tumor-adjacent normal tissues, but the level in epithelium was 5- to 10-fold lower. Analyzing DNA methylation of 25 CpG sites within 960 bp around the transcription initiation site of the SERPINE1 gene by bisulfite sequencing, we did the surprising observation that both tumors and tumor-adjacent normal tissue had a significant level of methylation, whereas there was very little methylation in tissue from healthy volunteers, suggesting that tumor-adjacent normal tissue already contains transformation-associated epigenetic changes. However, there was no general inverse correlation between PAI-1 mRNA levels and SERPINE1 gene methylation in all tissues, showing that CpG methylation is not the main determinant of the PAI-1 expression level in oral tissue.

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“…2 Generally, the invasive property of tumor cells is dependent on activation of proteases, such as plasmin at the cell surface. [3][4][5] Inactive plasminogen is converted to the active, broadspectrum serine proteinase, plasmin, by plasminogen activators (PA), including urokinase-type PA (uPA) and tissue-type PA (tPA), and their overexpression is considered central in malignant tumor progression. 6,7 Efficient plasmin generation involves plasminogen binding, together with PA, on cell surfaces by annexin A2, urokinasetype PA receptor, and other docking sites that colocalize enzyme and substrate.…”

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confidence: 99%

Secretory Leukocyte Protease Inhibitor (SLPI) Expression and Tumor Invasion in Oral Squamous Cell Carcinoma

Wen

Nikitakis

Chaisuparat

et al. 2011

The American Journal of Pathology

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Differential expression of secretory leukocyte protease inhibitor (SLPI) impacts on tumor progression. SLPI directly inhibits elastase and other serine proteases, and regulates matrix metalloproteinases, plasminogen activation, and plasmin downstream targets to influence invasion. We examined tissues from human oral squamous cell carcinoma (OSCC) for SLPI expression in parallel with proteases associated with tumor progression and evaluated their relationships using tumor cell lines. Significantly decreased SLPI was detected in OSCC compared to normal oral epithelium. Furthermore, an inverse correlation between SLPI and histological parameters associated with tumor progression, including stage of invasion, pattern of invasion, invasive cell grade, and composite histological tumor score was evident. Conversely, elevated plasmin and elastase were positively correlated with histological parameters of tumor invasion. In addition to its known inhibition of elastase, we identify SLPI as a novel inhibitor of plasminogen activation through its interaction with annexin A2 with concomitant reduced plasmin generation by macrophages and OSCC cell lines. In an in vitro assay measuring invasive activity, SLPI blocked protease-dependent tumor cell migration. Our data suggest that SLPI may possess antitumorigenic activity by virtue of its ability to interfere with multiple requisite proteolytic steps underlying tumor cell invasion and may provide insight into potential stratification of oral cancer according to risk of occult metastasis, guiding treatment strategies.

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Detection of plasminogen activators in oral cancer by laser capture microdissection combined with zymography

Cited by 20 publications

References 23 publications

Urinary-Type Plasminogen Activator Receptor/α3β1 Integrin Signaling, Altered Gene Expression, and Oral Tumor Progression

Urinary-Type Plasminogen Activator Receptor/α3β1 Integrin Signaling, Altered Gene Expression, and Oral Tumor Progression

Epigenetic alterations of the SERPINE1 gene in oral squamous cell carcinomas and normal oral mucosa

Secretory Leukocyte Protease Inhibitor (SLPI) Expression and Tumor Invasion in Oral Squamous Cell Carcinoma

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