Thrombin receptor overexpression in malignant and physiological invasion processes

Even-Ram, Sharona; Uziely, Beatrice; Cohen, Patrizia; Grisaru‐Granovsky, Sorina; Maoz, Myriam; Ginzburg, Yoav; Reich, Reuven; Vlodavsky, Israël; Bar-Shavit, Rachel

doi:10.1038/nm0898-909

Cited by 405 publications

(356 citation statements)

References 35 publications

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“…35 With this effect, our data demonstrate that thrombin as PAR1 agonist peptide stimulate invasion capacity of renal carcinoma cells. It has been proposed that VHL could play a role in invasion process.…”

Section: Discussionmentioning

confidence: 51%

“…ALEXA Fluor Rhodamine-phalloidin, ALEXA Fluor goat anti-mouse IgG and ProLong Antifade kit were purchased from Molecular Probes (Eugene, OR). 35 S-Met/Cys was obtained from ICN (Costa Mesa, CA). Protein G-Sepharose beads was purchased from Amersham Pharmacia Biotech (Uppsala, Sweden).…”

Section: Methodsmentioning

confidence: 99%

“…Then 100 Ci/ml of 35 S-Met/Cys and 1 U/ml of thrombin were added to the cells for 10 min. After labeling, cells were lysed in buffer A. Aliquots of lysates (250 g of protein) were precleared by incubation for 1 hr at 4°C with 20 l of protein G-Sepharose beads (50% in PBS).…”

Section: Metabolic Labeling and Immunoprecipitationmentioning

confidence: 99%

“…34,35 To determine the effect of PAR activation on the invasion capacity of Caki-1 cells, an in vitro assay through Matrigel was performed. Our results demonstrated that thrombin stimulated significantly Caki-1 invasion by 1.9-fold (Fig.…”

Section: Invasion Potential Of Caki-1 Cells Is Stimulated By Thrombinmentioning

confidence: 99%

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von Hippel‐Lindau tumor suppressor protein stimulation by thrombin involves RhoA activation

Turcotte¹,

Desrosiers²,

Brand³

et al. 2004

Intl Journal of Cancer

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Inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene is associated with the development of vascular tumors including renal cell carcinoma. Aside from the role played by the VHL protein (pVHL) in negative regulation of hypoxia-inducible factor, 41F-1␣, pVHL also takes part in cytoskeletal organization. Thrombin is a serine protease involved in angiogenesis and in cancer progression and its action is mediated by the protease-activated receptors (PARs). In several cell types, thrombin induces reorganization of the cytoskeleton along with RhoA activation. Thus, we conducted an investigation on the capacity of thrombin to regulate pVHL expression. Our results demonstrated that VHL mRNA and protein levels were increased by thrombin in cultured renal cancer cells. Cytoplasmic pVHL was redistributed to perinuclear regions and membrane fractions following thrombin treatments. Stimulation of Caki-1 cells with PAR1, PAR2 and PAR4 agonist peptides demonstrated that PAR1 was the receptor involved in thrombin-induced pVHL expression. Western blot analysis confirmed that these cells express PAR1 and that its expression was increased by thrombin. PAR1 activation by both thrombin and an agonist peptide stimulated renal cancer cell invasion through Matrigel. Interestingly, the upregulation of pVHL was dependent on RhoA because C3 exotoxin abolished pVHL induction. However, the pharmacological Rho kinase inhibitor, Y27632, did not influence pVHL expression in the presence of thrombin, suggesting that other RhoA effectors were involved in the process. Together, these results demonstrate that thrombin induces both pVHL expression via PAR1/RhoA activation as well as the stimulation of renal cancer cell invasion suggesting a role for thrombin in tumor invasion.

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Section: Discussionmentioning

confidence: 51%

Section: Methodsmentioning

confidence: 99%

Section: Metabolic Labeling and Immunoprecipitationmentioning

confidence: 99%

Section: Invasion Potential Of Caki-1 Cells Is Stimulated By Thrombinmentioning

confidence: 99%

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von Hippel‐Lindau tumor suppressor protein stimulation by thrombin involves RhoA activation

Turcotte¹,

Desrosiers²,

Brand³

et al. 2004

Intl Journal of Cancer

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“…7 PAR1 and PAR2 are important in tumor cell motility in melanoma 23,24 and PAR1 is involved in the invasive and metastatic processes of breast, 25,26 ovarian, 6 and pancreatic cancer. 27 PAR1 also has been identified as an oncogene in the transformation of NIH3T3 mouse fibroblasts 28,29 and ectopic expression of PAR1 confers tumorigenicity in PAR1-null breast carcinoma cells.…”

mentioning

confidence: 99%

Targeting Protease-Activated Receptor-1 with Cell-Penetrating Pepducins in Lung Cancer

Cisowski

O’Callaghan

Kuliopulos

et al. 2011

The American Journal of Pathology

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Protease-activated receptors (PARs) are G-proteincoupled receptors that are activated by proteolytic cleavage and generation of a tethered ligand. High PAR1 expression has been documented in a variety of invasive cancers of epithelial origin. In the present study, we investigated the contribution of the four PAR family members to motility of lung carcinomas and primary tumor samples from patients. We found that of the four PARs, only PAR1 expression was highly increased in the lung cancer cell lines. Primary lung cancer cells isolated from patient lung tumors migrated at a 10-to 40-fold higher rate than epithelial cells isolated from nonmalignant lung tissue. Cellpenetrating pepducin inhibitors were generated against the first (i1) and third (i3) intracellular loops of PAR1 and tested for their ability to inhibit PAR1-driven migration and extracellular regulated kinase (ERK)1/2 activity. The PAR1 pepducins showed significant inhibition of cell migration in both primary and established cell lines similar to silencing of PAR1 expression with short hairpin RNA (shRNA). Unlike i1 pepducins, the i3 loop pepducins were effective inhibitors of PAR1-mediated ERK activation and tumor growth. Comparable in efficacy with Bevacizumab, monotherapy with the PAR1 i3 loop pepducin P1pal-7 provided significant 75% inhibition of lung tumor growth in nude mice. We identify the PAR1-ERK1/2 pathway as a feasible target for therapy in lung cancer. Lung cancer is the leading cause of cancer deaths in the United States and worldwide, and is the second most common cancer overall. 1 The majority of patients eventually develop distant metastases, which leads to substantial morbidity and mortality. Currently available chemotherapeutic regimens for the treatment of non-small-cell lung cancer (NSCLC) include combinations of cisplatin or carboplatin, and etoposide, paclitaxel, docetaxel, gemcitabine, vinorelbin, and irinotecan. These regimens are generally not curative and may confer modest prolongation of life and symptomatic relief. 2,3 More recently, targeted therapies have become available for the treatment of lung cancer. These include small molecules and antibodies that target epidermal growth factor receptor and vascular endothelial growth factor receptor. However, the currently available molecular therapies still result in relatively modest prolongation of median and overall survival, pointing to the necessity for developing more effective treatment modalities for patients with advanced NSCLC.Emerging evidence has identified protease activated receptor-1 (PAR1) as a promising target to impact tumor progression, metastasis, and angiogenesis in a variety of cancers including breast, ovarian, melanoma, prostate, and colon cancer. 4 -7 However, the role of PAR1 and the other PAR family members in lung cancer is largely unexplored. To date, four different PARs have been identified: PAR1, PAR2, PAR3, and PAR4. 8,9 -13 PAR1 originally was discovered on platelets and serves as the prototype for this specialized class of proteolytically activ...

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Vascular protease receptors: integrating haemostasis and endothelial cell functions

2000

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The endothelium plays a crucial dynamic role as a protective interface between blood and the underlying tissues during the haemostatic process, which maintains blood flow in the circulation and prevents life‐threatening blood loss. Following vessel wall injury with initial platelet adhesion and aggregation to exposed subendothelial extracellular matrix, the initiation, amplification, and control of haemostasis depend on structurally unrelated membrane‐associated receptors for blood coagulation proteases including tissue factor, G‐protein‐coupled protease‐activatable receptors, thrombomodulin, and protein C receptor, respectively. In addition to their regulatory role in haemostasis, the respective (pro‐)enzyme ligands such as Factors VIIa and Xa, thrombin or protein C mediate specific signalling pathways in vascular cells related to migration, proliferation or adhesion. The functional importance of these receptors beyond haemostasis has been manifested by various lethal and pathological phenotypes in knock‐out mice. These protease receptors thereby provide important molecular links in the vascular system and serve to integrate haemostasis with endothelial cell functions which are relevant for the (patho‐)physiological responses to injury or inflammatory challenges. Copyright © 2000 John Wiley & Sons, Ltd.

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Thrombin receptor overexpression in malignant and physiological invasion processes

Cited by 405 publications

References 35 publications

von Hippel‐Lindau tumor suppressor protein stimulation by thrombin involves RhoA activation

von Hippel‐Lindau tumor suppressor protein stimulation by thrombin involves RhoA activation

Targeting Protease-Activated Receptor-1 with Cell-Penetrating Pepducins in Lung Cancer

Vascular protease receptors: integrating haemostasis and endothelial cell functions

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