The role of vascular endothelial growth factor and matrix metalloproteinases in canine lymphoma: in vivo and in vitro study

BMC Veterinary Research

Luca Aresu³  Massimo Castagnaro³  Mauro Dacasto³  Sabine E Essler¹  Barbara C Rütgen⁵  Stefano Comazzi²  Fulvio Riondato⁴  Maria Elena Gelain³  Mery Giantin³  Arianna Aricò³ 

[1] Institute of Immunology, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterinärplatz 1, Vienna, 1210, Austria;Department of Animal Pathology, Public Health and Veterinary Hygiene, Faculty of Veterinary Medicine, University of Milano, Via Celoria 10, Milan, 20133, Italy;Department of Comparative Biomedicine and Food Science, University of Padova, Viale dell'Università 16, Agripolis Legnaro, PD, 35020, Italy;Department of Animal Pathology, Faculty of Veterinary Medicine, University of Torino, Via Leonardo da Vinci 44, Grugliasco, TO, 10095, Italy;Clinical Pathology, Department of Pathobiology, University of Veterinary Medicine Vienna, Veterinärplatz 1, Vienna, 1210, Austria

关键词: VEGF;    TIMPs;    MMPs;    Lymphoma;    Phenotype;    Dog;   

Others  :  1119544
DOI  :  10.1186/1746-6148-9-94

 received in 2012-10-25, accepted in 2013-05-01,  发布年份 2013

【 摘 要 】

Background

Canine lymphoma represents the most frequent haematopoietic cancer and it shares some similarities with human non-Hodgkin lymphoma. Matrix metalloproteinases (MMPs) and vascular endothelial growth factor (VEGF) play a coordinated role during invasion and proliferation of malignant cells; however, little is known about their role in canine haematologic malignancies. The aim of this study was to investigate the mRNA and protein expression of VEGF and the most relevant MMPs in canine lymphoma. Lymph node aspirates from 26 B-cell and 21 T-cell lymphomas were collected. The protein expression levels of MMP-9, MMP-2 and VEGF-A were evaluated by immunocytochemistry, and the mRNA levels of MMP-2, MMP-9, MT1-MMP, TIMP-1, TIMP-2, RECK, VEGF-A and VEGF-164 were measured using quantitative RT-PCR.

Results

MT1-MMP, TIMP-1 and RECK mRNA levels were significantly higher in T-cell lymphomas than in B-cell lymphomas. Higher mRNA and protein levels of MMP-9 and VEGF-A were observed in T-cell lymphomas than in B-cell lymphomas and healthy control lymph nodes. A positive correlation was found between MMP-9 and VEGF-A in T-cell lymphomas. Moreover, MMP-9, MT1-MMP, TIMP-1 and VEGF-A were expressed at the highest levels in high-grade T-cell lymphomas.

Conclusions

This study provides new information on the expression of different MMPs and VEGF in canine lymphoma, suggesting a possible correlation between different MMPs and VEGF, immunophenotype and prognosis.

【 授权许可】

   
2013 Aricò et al.; licensee BioMed Central Ltd.

【 图 表 】

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【 参考文献 】

• [1]Breen M, Modiano JF: Evolutionarily conserved cytogenetic changes in hematological malignancies of dogs and humans-man and his best friend share more than companionship. Chromosome Res 2008, 16(1):145-154.
• [2]Marconato L, Gelain ME, Comazzi S: The dog as a possible animal model for human non-Hodgkin lymphoma: a review. Hematol Oncol 2013, 31(1):1-9.
• [3]Pennanen H, Kuittinen O, Soini Y, Turpeenniemi-Hujanen T: Prognostic significance of p53 and matrix metalloproteinase-9 expression in follicular lymphoma. Eur J Haematol 2008, 81(4):289-97.
• [4]Kossakowska AE, Urbanski SJ, Janowska-Wieczorek A: Matrix metalloproteinases and their tissue inhibitors - expression, role and regulation in human malignant non-Hodgkin's lymphomas. Leuk Lymphoma 2000, 39(5–6):485-93.
• [5]Egeblad M, Werb Z: New functions for the matrix metalloproteinases in cancer progression. Nat Rev Cancer 2002, 2:161-74.
• [6]Aresu L, Giantin M, Morello E, Vascellari M, Castagnaro M, Lopparelli R, Zancanella V, Granato A, Garbisa S, Aricò A, Bradaschia A, Mutinelli F, Dacasto M: Matrix metalloproteinases and their inhibitors in canine mammary tumors. BMC Vet Res 2011, 7:33. BioMed Central Full Text
• [7]Kurschat P, Zigrino P, Nischt R, Breitkopf K, Steurer P, Klein CE, Krieg T, Mauch C: Tissue inhibitor of matrix metalloproteinase-2 regulates matrix metalloproteinase-2 activation by modulation of membrane-type 1 matrix metalloproteinase activity in high and low invasive melanoma cell lines. J Biol Chem 1999, 274(30):21056-62.
• [8]Takagi S, Kato Y, Asano K, Ohsaki T, Bosnakovski D, Hoshino Y, Okumura M, Kadosawa T, Fujinaga T: Matrix metalloproteinase inhibitor RECK expression in canine tumors. J Vet Med Sci 2005, 67(8):761-7.
• [9]Bergers G, Brekken R, McMahon G, Vu HT, Itoh T, Tamaki K, Tanzawa K, Thorpe P, Itohara S, Werb Z, Hanahan D: Matrix metalloproteinase-9 triggers the angiogenic switch during carcinogenesis. Nat Cell Biol 2000, 2(10):737-44.
• [10]Giantin M, Aresu L, Benali S, Aricò A, Morello EM, Martano M, Vascellari M, Castagnaro M, Lopparelli RM, Zancanella V, Granato A, Mutinelli F, Dacasto M: Expression of Matrix Metalloproteinases, Tissue Inhibitors of Metalloproteinases and Vascular Endothelial Growth Factor in Canine Mast Cell Tumours. J Comp Pathol 2012, 147(4):419-29.
• [11]Göritz M, Müller K, Krastel D, Staudacher G, Schmidt P, Kühn M, Nickel R, Schoon HA: Canine Splenic Haemangiosarcoma: Influence of Metastases, Chemotherapy and Growth Pattern on Post-splenectomy Survival and Expression of Angiogenic Factors. J Comp Pathol 2012. [Epub ahead of print]
• [12]Aricò A, Giantin M, Gelain M, Riondato F, Mortarino M, Comazzi S, Dacasto M, Castagnaro M, Aresu L: Matrix metalloproteinases and vascular endothelial growth factor expression in canine leukaemias. Vet J 2012. [Epub ahead of print]
• [13]Aresu L, Aricò A, Comazzi S, Gelain ME, Riondato F, Mortarino M, Morello M, Stefanello D, Castagnaro M: VEGF and MMP-9: biomarkers for canine lymphoma. Vet Comp Oncol 2012. [Epub ahead of print]
• [14]Gentilini F, Calzolari C, Turba ME, Agnoli C, Fava D, Forni M, Bergamini PF: Prognostic value of serum vascular endothelial growth factor (VEGF) and plasma activity of matrix metalloproteinase (MMP) 2 and 9 in lymphoma-affected dog. Leuk Res 2005, 29(11):1263-9.
• [15]Wolfesberger B, Guija De Arespacohaga A, Willmann M, Gerner W, Miller I, Schwendenwein I, Kleiter M, Egerbacher M, Thalhammer JG, Muellauer L, Skalicky M, Walter I: Expression of vascular endothelial growth factor and its receptors in canine lymphoma. J Comp Pathol 2007, 137(1):30-40.
• [16]Zizzo N, Patruno R, Zito FA, Di Summa A, Tinelli A, Troilo S, Misino A, Ruggieri E, Goffredo V, Gadaleta CD, Ranieri G: Vascular endothelial growth factor concentrations from platelets correlate with tumor angiogenesis and grading in a spontaneous canine non-Hodgkin lymphoma model. Leuk Lymphoma 2010, 51(2):291-6.
• [17]Fournel-Fleury C, Magnol JP, Bricaire P, Marchal T, Chabanne L, Delverdier A, Bryon PA, Felman P: Cytohistological and immunological classification of canine malignant lymphomas: comparison with human non hodgkin’s lymphomas. J Comp Pathol 1997, 117(1):55-59.
• [18]Rütgen BC, Hammer SE, Gerner W, Christian M, de Arespacochaga AG, Willmann M, Kleiter M, Schwendenwein I, Saalmüller A: Establishment and characterization of a novel canine B-cell line derived from a spontaneously occurring diffuse large cell lymphoma. Leuk Res 2010, 34(7):932-8.
• [19]Kisseberth WC, Nadella MV, Breen M, Thomas R, Duke SE, Murahari S, Kosarek CE, Vernau W, Avery AC, Burkhard MJ, Rosol TJ: A novel canine lymphoma cell line: a translational and comparative model for lymphoma research. Leuk Res 2007, 31(12):1709-20.
• [20]Dunmire V, Wu C, Symmans WF, Zhang W: Increased yield of total RNA from fine-needle aspirates for use in expression microarray analysis. Biotechniques 2002, 33(4):890-2. 894, 896
• [21]Usui T, Ishida S, Yamashiro K, Kaji Y, Poulaki V, Moore J, Moore T, Amano S, Horikawa Y, Dartt D, Golding M, Shima DT, Adamis AP: VEGF164(165) as the pathological isoform: differential leukocyte and endothelial responses through VEGFR1 and VEGFR2. Invest Ophthalmol Vis Sci 2004, 45(2):368-74.
• [22]Lee S, Jo M, Koh SS, Kim S: Identification of novel universal housekeeping genes by statistical analysis of microarray data. J Biochem Mol Biol 2007, 40(2):226-231.
• [23]Livak KJ, Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)). Method 2001, 25(4):402-408.
• [24]Davies B, Waxman J, Wasan H, Abel P, Williams G, Krausz T, Neal D, Thomas D, Hanby A, Balkwill F: Levels of matrix metalloproteases in bladder cancer correlate with tumor grade and invasion. Cancer Res 1993, 53(22):5365-9.
• [25]Hayashibara T, Yamada Y, Onimaru Y, Tsutsumi C, Nakayama S, Mori N, Miyanishi T, Kamihira S, Tomonaga M, Maita T: Matrix metalloproteinase-9 and vascular endothelial growth factor: a possible link in adult T-cell leukaemia cell invasion. Br J Haematol 2002, 116(1):94-102.
• [26]Pedersen LM, Klausen TW, Davidsen UH, Johnsen HE: Early changes in serum IL-6 and VEGF levels predict clinical outcome following first-line therapy in aggressive non-Hodgkin's Lymphoma. Ann Hematol 2005, 84(8):510-516.
• [27]Buggins AG, Levi A, Gohil S, Fishlock K, Patten PE, Calle Y, Yallop D, Devereux S: Evidence for a macromolecular complex in poor prognosis CLL that contains CD38, CD49d, CD44 and MMP-9. Br J Haematol 2011, 154(2):216-22.
• [28]Vacca A, Ribatti D, Ria R, Pellegrino A, Bruno M, Merchionne F, Dammacco F: Proteolytic activity of human lymphoid tumor cells, Correlation with tumor progression. Dev Immunol 2000, 7(2–4):77-88.
• [29]Moehler TM, Ho AD, Goldschmidt H, Barlogie B: Angiogenesis in hematologic malignancies. Crit Rev Oncol Hematol 2003, 45(3):227-44.
• [30]Kossakowska AE, Urbanski SJ, Edwards DR: Tissue inhibitor of metalloproteinases-1 (TIMP-1) RNA is expressed at elevated levels in malignant non-Hodgkin's lymphomas. Blood 1991, 77(11):2475-81.
• [31]Kossakowska AE, Edwards DR, Prusinkiewicz C, Zhang MC, Guo D, Urbanski SJ, Grogan T, Marquez LA, Janowska-Wieczorek A: Interleukin-6 regulation of matrix metalloproteinase (MMP-2 and MMP-9) and tissue inhibitor of metalloproteinase (TIMP-1) expression in malignant non-Hodgkin's lymphomas. Blood 1999, 94(6):2080-9.
• [32]Caenazzo C, Onisto M, Sartor L, Scalerta R, Giraldo A, Nitti D, Garbisa S: Augmented membrane type 1 matrix metalloproteinase (MT1-MMP):MMP-2 messenger RNA ratio in gastric carcinomas with poor prognosis. Clin Cancer Res 1998, 4(9):2179-86.
• [33]Shofuda K, Moriyama K, Nishihashi A, Higashi S, Mizushima H, Yasumitsu H, Miki K, Sato H, Seiki M, Miyazaki K: Role of tissue inhibitor of metalloproteinases-2 (TIMP-2) in regulation of pro-gelatinase A activation catalyzed by membrane-type matrix metalloproteinase-1 (MT1-MMP) in human cancer cells. J Biochem 1998, 124(2):462-70.
• [34]Snoek-van Beurden PA, Von den Hoff JW: Zymographic techniques for the analysis of matrix metalloproteinases and their inhibitors. Biotechniques 2005, 38(1):73-83.
• [35]Wolfesberger B, Tonar Z, Fuchs-Baumgartinger A, Walter I, Skalicky M, Witter K, Thalhammer JG, Pagitz M, Kleiter M: Angiogenic markers in canine lymphoma tissues do not predict survival times in chemotherapy treated dogs. Res Vet Sci 2012, 92(3):444-50.