Expression of tissue factor signaling pathway elements correlates with the production of vascular endothelial growth factor and interleukin-8 in human astrocytoma patients

Carneiro-Lobo,Tatiana C.; Lima,Marina T.; Mariano‑Oliveira,Andréa; Dutra‑Oliveira,Angélica; Oba-Shinjo,Sueli M.; Marie,Suely K.N.; Sogayar,Mari C.; Monteiro,Robson Q.

doi:10.3892/or.2013.2880

Expression of tissue factor signaling pathway elements correlates with the production of vascular endothelial growth factor and interleukin-8 in human astrocytoma patients

Authors:
- Tatiana C. Carneiro-Lobo
- Marina T. Lima
- Andréa Mariano‑Oliveira
- Angélica Dutra‑Oliveira
- Sueli M. Oba-Shinjo
- Suely K.N. Marie
- Mari C. Sogayar
- Robson Q. Monteiro
View Affiliations

Affiliations: Institute of Medical Biochemistry, Federal University of Rio de Janeiro, RJ, Brazil, Biochemistry Department, Chemistry Institute, Cell and Molecular Therapy Center (NUCEL), University of São Paulo, SP, Brazil, Department of Neurology, School of Medicine, University of São Paulo, SP, Brazil
Published online on: November 28, 2013 https://doi.org/10.3892/or.2013.2880
Pages: 679-686

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

The expression levels of tissue factor (TF), the clotting initiator protein, have been correlated with angiogenesis and the histological grade of malignancy in glioma patients. The pro-tumor function of TF is linked to a family of G protein-coupled receptors known as protease-activated receptors (PARs), which may be activated by blood coagulation proteases. Activation of PARs elicits a number of responses, including the expression of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8). In the present study, we analyzed the expression of TF signaling pathway elements (TF, PAR1 and PAR2) and evaluated their correlation with the expression of downstream products (VEGF and IL-8) in human astrocytoma patients. Quantitative PCR (qPCR) showed a significant increase in TF expression in grade IV (glioblastoma) tumors, which was inversely correlated with the expression of the tumor-suppressor PTEN. Immunohistochemistry and qPCR analyses demonstrated a highly significant elevation in the expression of PAR1, but not PAR2, in tumor samples from high-grade astrocytoma patients. The elevated VEGF expression levels detected in the high-grade astrocytoma samples were positively correlated with TF, PAR1 and PAR2 expression. In addition, IL-8 was significantly increased in glioblastoma patients and positively correlated with TF and PAR2 expression. Further in vitro assays employing the human glioma cell lines U87-MG and HOG demonstrated that a synthetic peptide PAR2 agonist stimulated VEGF and IL-8 production. Our findings suggest a role for TF signaling pathway elements in astrocytoma progression, particularly in glioblastoma. Therefore, TF/PAR signaling elements may be suitable targets for the development of new therapies for the treatment of aggressive glioma.

View Figures

View References

1	Louis DN, Ohgaki H, Wiestler OD, Cavenee WK, Burger PC, Jouvet A, Scheithauer BW and Kleihues P: The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol. 114:97–109. 2007. View Article : Google Scholar : PubMed/NCBI
2	Wen PY and Kesari S: Malignant gliomas in adults. N Engl J Med. 359:492–507. 2008. View Article : Google Scholar : PubMed/NCBI
3	Williams JC and Mackman N: Tissue factor in health and disease. Front Biosci. 1:358–372. 2012. View Article : Google Scholar
4	Francischetti IM, Seydel KB and Monteiro RQ: Blood coagulation, inflammation, and malaria. Microcirculation. 15:81–107. 2008. View Article : Google Scholar
5	Rak J, Milsom C, Magnus N and Yu J: Tissue factor in tumour progression. Best Pract Res Clin Haematol. 22:71–83. 2009. View Article : Google Scholar : PubMed/NCBI
6	Ruf W, Disse J, Carneiro-Lobo TC, Yokota N and Schaffner F: Tissue factor and cell signalling in cancer progression and thrombosis. J Thromb Haemost. 9(Suppl 1): 306–315. 2011. View Article : Google Scholar : PubMed/NCBI
7	Lima LG and Monteiro RQ: Activation of blood coagulation in cancer: implications for tumor progression. Biosci Rep. 33:e000642013.PubMed/NCBI
8	Tehrani M, Friedman TM, Olson JJ and Brat DJ: Intravascular thrombosis in central nervous system malignancies: a potential role in astrocytoma progression to glioblastoma. Brain Pathol. 18:164–171. 2008. View Article : Google Scholar : PubMed/NCBI
9	Brat DJ and Van Meir EG: Vaso-occlusive and prothrombotic mechanisms associated with tumor hypoxia, necrosis, and accelerated growth in glioblastoma. Lab Invest. 84:397–405. 2004. View Article : Google Scholar : PubMed/NCBI
10	Anand M and Brat DJ: Oncogenic regulation of tissue factor and thrombosis in cancer. Thromb Res. 129(Suppl 1): S46–S49. 2012. View Article : Google Scholar : PubMed/NCBI
11	Rong Y, Post DE, Pieper RO, Durden DL, Van Meir EG and Brat DJ: PTEN and hypoxia regulate tissue factor expression and plasma coagulation by glioblastoma. Cancer Res. 65:1406–1413. 2005. View Article : Google Scholar : PubMed/NCBI
12	Kasthuri RS, Taubman MB and Mackman N: Role of tissue factor in cancer. J Clin Oncol. 27:4834–4838. 2009. View Article : Google Scholar : PubMed/NCBI
13	Coughlin SR: Protease-activated receptors in hemostasis, thrombosis and vascular biology. J Thromb Haemos. 3:1800–1814. 2005. View Article : Google Scholar : PubMed/NCBI
14	Elste AP and Petersen I: Expression of proteinase-activated receptor 1–4 (PAR 1–4) in human cancer. J Mol Histol. 41:89–99. 2010.
15	Albrektsen T, Sørensen BB, Hjortø GM, Fleckner J, Rao LV and Petersen LC: Transcriptional program induced by factor VIIa-tissue factor, PAR1 and PAR2 in MDA-MB-231 cells. J Thromb Haemost. 5:1588–1597. 2007. View Article : Google Scholar : PubMed/NCBI
16	Gessler F, Voss V, Dützmann S, Seifert V, Gerlach R and Kögel D: Inhibition of tissue factor/protease-activated receptor-2 signaling limits proliferation, migration and invasion of malignant glioma cells. Neuroscience. 165:1312–1322. 2010. View Article : Google Scholar : PubMed/NCBI
17	Oba-Shinjo SM, Bengtson MH, Winnischofer SM, Colin C, Vedoy CG, de Mendonça Z, Marie SK and Sogayar MC: Identification of novel differentially expressed genes in human astrocytomas by cDNA representational difference analysis. Brain Res Mol Brain Res. 140:25–33. 2005. View Article : Google Scholar : PubMed/NCBI
18	Carneiro-Lobo TC, Konig S, Machado DE, Nasciutti LE, Forni MF, Francischetti IM, Sogayar MC and Monteiro RQ: Ixolaris, a tissue factor inhibitor, blocks primary tumor growth and angiogenesis in a glioblastoma model. J Thromb Haemost. 7:1855–1864. 2009. View Article : Google Scholar : PubMed/NCBI
19	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCt. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
20	De Groot CJ, Langeveld CH, Jongenelen CA, Montagne L, Van Der Valk P and Dijkstra CD: Establishment of human adult astrocyte cultures derived from postmortem multiple sclerosis and control brain and spinal cord regions: immunophenotypical and functional characterization. J Neurosci Res. 49:342–354. 1997.
21	Dutra-Oliveira A, Monteiro RQ and Mariano-Oliveira A: Protease-activated receptor-2 (PAR2) mediates VEGF production through the ERK1/2 pathway in human glioblastoma cell lines. Biochem Biophys Res Commun. 421:221–227. 2012. View Article : Google Scholar : PubMed/NCBI
22	Hamada K, Kuratsu J, Saitoh Y, Takeshima H, Nishi T and Ushio Y: Expression of tissue factor correlates with grade of malignancy in human glioma. Cancer. 77:1877–1883. 1996. View Article : Google Scholar : PubMed/NCBI
23	Guan M, Jin J, Su B, Liu WW and Lu Y: Tissue factor expression and angiogenesis in human glioma. Clin Biochem. 35:321–325. 2002. View Article : Google Scholar : PubMed/NCBI
24	Plate KH, Breier G, Weich HA and Risau W: Vascular endothelial growth factor is a potential tumour angiogenesis factor in human gliomas in vivo. Nature. 359:845–848. 1992. View Article : Google Scholar : PubMed/NCBI
25	Koch AE, Polverini PJ, Kunkel SL, Harlow LA, DiPietro LA, Elner VM, Elner SG and Strieter RM: Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science. 258:1798–1801. 1992. View Article : Google Scholar : PubMed/NCBI
26	Brat DJ, Bellail AV and Van Meir EG: The role of interleukin-8 and its receptors in gliomagenesis and tumoral angiogenesis. Neuro Oncol. 7:122–133. 2005. View Article : Google Scholar : PubMed/NCBI
27	Magnus N, Garnier D and Rak J: Oncogenic epidermal growth factor receptor up-regulates multiple elements of the tissue factor signaling pathway in human glioma cells. Blood. 116:815–818. 2010. View Article : Google Scholar : PubMed/NCBI
28	Lima FR, Kahn SA, Soletti RC, Biasoli D, Alves T, da Fonseca AC, Garcia C, Romão J, Brito L, Holanda-Afonso R, Faria J, Borges H and Moura-Neto V: Glioblastoma: therapeutic challenges, what lies ahead. Biochim Biophys Acta. 1826:338–349. 2012.PubMed/NCBI
29	Fernandes RS, Kirszberg C, Rumjanek VM and Monteiro RQ: On the molecular mechanisms for the highly procoagulant pattern of C6 glioma cells. J Thromb Haemost. 4:1546–1552. 2006. View Article : Google Scholar : PubMed/NCBI
30	Kirszberg C, Lima LG, Da Silva de Oliveira A, Pickering W, Gray E, Barrowcliffe TW, Rumjanek VM and Monteiro RQ: Simultaneous tissue factor expression and phosphatidylserine exposure account for the highly procoagulant pattern of melanoma cell lines. Melanoma Res. 19:301–308. 2009. View Article : Google Scholar : PubMed/NCBI
31	Zhang Y, Zhan H, Xu W, Yuan Z, Lu P, Zhan L and Li Q: Upregulation of matrix metalloproteinase-1 and proteinase-activated receptor-1 promotes the progression of human gliomas. Pathol Res Pract. 207:24–29. 2011. View Article : Google Scholar : PubMed/NCBI
32	Boire A, Covic L, Agarwal A, Jacques S, Sherifi S and Kuliopulos A: PAR1 is a matrix metalloprotease-1 receptor that promotes invasion and tumorigenesis of breast cancer cells. Cell. 120:303–313. 2005. View Article : Google Scholar : PubMed/NCBI
33	Yin YJ, Salah Z, Maoz M, Even Ram SC, Ochayon S, Neufeld G, Katzav S and Bar-Shavit R: Oncogenic transformation induces tumor angiogenesis: a role for PAR1 activation. FASEB J. 17:163–174. 2003. View Article : Google Scholar : PubMed/NCBI
34	Xu Y, Gu Y, Keep RF, Heth J, Muraszko KM, Xi G and Hua Y: Thrombin up-regulates vascular endothelial growth factor in experimental gliomas. Neurol Res. 31:759–765. 2009. View Article : Google Scholar : PubMed/NCBI
35	Versteeg HH, Schaffner F, Kerver M, Ellies LG, Andrade-Gordon P, Mueller BM and Ruf W: Protease-activated receptor (PAR) 2, but not PAR1, signaling promotes the development of mammary adenocarcinoma in polyoma middle T mice. Cancer Res. 68:7219–7227. 2008. View Article : Google Scholar : PubMed/NCBI
36	Schaffner F, Versteeg HH, Schillert A, Yokota N, Petersen LC, Mueller BM and Ruf W: Cooperation of tissue factor cytoplasmic domain and PAR2 signaling in breast cancer development. Blood. 116:6106–6113. 2010. View Article : Google Scholar : PubMed/NCBI
37	Svensson KJ, Kucharzewska P, Christianson HC, Sköld S, Löfstedt T, Johansson MC, Mörgelin M, Bengzon J, Ruf W and Belting M: Hypoxia triggers a proangiogenic pathway involving cancer cell microvesicles and PAR-2-mediated heparin-binding EGF signaling in endothelial cells. Proc Natl Acad Sci USA. 108:13147–13152. 2011. View Article : Google Scholar : PubMed/NCBI
38	Harter PN, Dützmann S, Drott U, Zachskorn C, Hattingen E, Capper D, Gessler F, Senft C, Seifert V, Plate KH, Kögel D and Mittelbronn M: Anti-tissue factor (TF9–10H10) treatment reduces tumor cell invasiveness in a novel migratory glioma model. Neuropathology. 33:515–525. 2013.
39	Ornstein DL, Meehan KR and Zacharski LR: The coagulation system as a target for the treatment of human gliomas. Semin Thromb Hemost. 28:19–28. 2002. View Article : Google Scholar : PubMed/NCBI
40	Hua Y, Tang L, Keep RF, Schallert T, Fewel ME, Muraszko KM, Hoff JT and Xi G: The role of thrombin in gliomas. J Thromb Haemost. 3:1917–1923. 2005. View Article : Google Scholar : PubMed/NCBI
41	Carneiro-Lobo TC, Schaffner F, Disse J, Ostergaard H, Francischetti IM, Monteiro RQ and Ruf W: The tick-derived inhibitor Ixolaris prevents tissue factor signaling on tumor cells. J Thromb Haemost. 10:1849–1858. 2012. View Article : Google Scholar : PubMed/NCBI