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Evaluation of the expression of c-kit, PTEN, and correlation among connexins 32, 43, e-cadherin and cadherin-associated proteins in canine mast cell tumors

Abstract

The study of mast cell tumors is very important to understand their underlying causes, types and treatment modalities. The subject is very important because the tumor is very frequent in dogs. Mast cell tumors originate in the dermis and represent 16 to 21% of all cutaneous tumors. The biological behaviour of the mast cell tumors is variable, and therefore it is difficult to perform diagnosis and therapy. Neoplastic cells exhibit variable degrees of differentiation, based on the presence and appearance of cytoplasmic granules in the histological sections stained with hematoxylin and eosin. Canine mast cell tumors were classified in degrees \I, II or III, according to Patnaik et al., 1984. This classification is being used subjectively in these tumors to establish the prognosis. The aim of this study is to study the molecular basis of the cell-cell adhesion genes and of the communicating junctions. Our specific objectives are: 1) Perform histopathologic analysis of the mast cell tumors and establish the classification according to Patnaik et al., 1984; 2) To establish and characterize the primary culture of canine mast cell tumors grades I, II or III; 3) Perform immunohistochemical analysis of the proteins c-kit and PTEN (phosphatase, tensin homologue) and of the following adhesion molecules: E-cadherin, ²-catenin, ±-catenin, p120-catenin and cell communication molecules like connexins 32 and 43, in the different tumor grades; 4) Isolate the mast cells from tumor cultures using magnetic columns, in order to obtain proteins from mast cells only to the analysis of expression of the molecules c-kit, PTEN, Cx32, Cx43, E-cadherin, ²-catenin, ±-catenin, p120-catenin by immunoblot; 5)To extract total RNA from mast cells to the analysis of CD117 (c-kit), Phosphatase, tensin homologue (PTEN), GJB1 (Cx32), GJA1 (Cx43), CDH1 (E-caderina), CTNNB1 (²-catenina), CTNNA1 (±-catenina) and CTNND1 (p120-catenin), by Real-Time PCR. (AU)

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Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
MACKOWIAK, I. I.; GENTILE, L. B.; CHAIBLE, L. M.; NAGAMINE, M. K.; GUERRA, J. M.; MOTA, E. F. F.; MATERA, J. M.; MENNECIER, G.; SANCHES, D. S.; DAGLI, M. L. Z. E-cadherin in canine mast cell tumors: Decreased expression and altered subcellular localization in Grade 3 tumors. VETERINARY JOURNAL, v. 194, n. 3, p. 405-411, DEC 2012. Web of Science Citations: 1.

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