Snai-1 and Epithelial-Mesenchymal Transition-Related Protein Immunoexpression in Canine Mammary Carcinomas
Author(s)
Breno S. Salgado1*,
Rafael M. Rocha3,
Fernando A. Soares3,
Fátima Gärtner4,
Noeme S. Rocha1,2
Affiliation(s)
1 Departamento de Patologia, Faculdade de Medicina de Botucatu, Universidade Estadual Paulista (UNESP), Botucatu, Brazil. 2 Laboratório de Patologia Investigativa e Comparada, Departamento de Clínica Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista (UNESP), Botucatu, Brazil. 3 Departamento de Anatomia Patológica, Hospital A. C. Camargo, Fundacao Antonio Prudente, Sao Paulo, Brazil. 4 Institute of Pathology and Molecular Immunology (IPATIMUP), Oporto, Portugal.
1 Departamento de Patologia, Faculdade de Medicina de Botucatu, Universidade Estadual Paulista (UNESP), Botucatu, Brazil. 2 Laboratório de Patologia Investigativa e Comparada, Departamento de Clínica Veterinária, Faculdade de Medicina Veterinária e Zootecnia, Universidade Estadual Paulista (UNESP), Botucatu, Brazil. 3 Departamento de Anatomia Patológica, Hospital A. C. Camargo, Fundacao Antonio Prudente, Sao Paulo, Brazil. 4 Institute of Pathology and Molecular Immunology (IPATIMUP), Oporto, Portugal.
ABSTRACT
Epithelial-mesenchymal transition (EMT) is defined as switching of polarized epithelial cells to a migratory fibroblastoid phenotype. EMT is known to be involved in the progression and metastasis of various cancers in humans, but this specific process is still little explored in the veterinary literature. The aim of this research was to evaluate the expression of EMT-related proteins in canine mammary carcinomas (CMCs). The expression of six EMT-related proteins in 94 CMCs of female dogs was evaluated by immunohistochemistry using a tissue array method. Additionally, clinicopathological characteristics were compared with the expression of EMT-related proteins. Loss of epithelial protein and/or acquisition of the expression of mesenchymal proteins were observed in CMCs. Loss of epithelial protein and/or acquisition of the expression of mesenchymal proteins were observed, particularly in tumors with evidence of stromal invasion; however, significance was only observed between the S100A4 and vascular invasion. In addition, Snai-1 nuclear immunoexpression was significantly related to E-cadherin loss. In conclusion, loss of epithelial proteins and/or the acquisition of mesenchymal proteins are associated with EMT and may have an important role in the evaluation of CMC patients. The unique immunoexpression pattern of Snai-1 could help to distinguish between an adenoma and a non-metastatic carcinoma and seems to be related to conversion of myoepithelial cells to a complete mesenchymal-like phenotype. Loss of E-cadherin and cytokeratin and change of immunoexpression pattern of Snai-1, N-cadherin, S100A4 and MMP-2 indicate the occurrence of EMT in canine mammary carcinomas and should result in an en bloc resection or a close follow-up.
Epithelial-mesenchymal transition (EMT) is defined as switching of polarized epithelial cells to a migratory fibroblastoid phenotype. EMT is known to be involved in the progression and metastasis of various cancers in humans, but this specific process is still little explored in the veterinary literature. The aim of this research was to evaluate the expression of EMT-related proteins in canine mammary carcinomas (CMCs). The expression of six EMT-related proteins in 94 CMCs of female dogs was evaluated by immunohistochemistry using a tissue array method. Additionally, clinicopathological characteristics were compared with the expression of EMT-related proteins. Loss of epithelial protein and/or acquisition of the expression of mesenchymal proteins were observed in CMCs. Loss of epithelial protein and/or acquisition of the expression of mesenchymal proteins were observed, particularly in tumors with evidence of stromal invasion; however, significance was only observed between the S100A4 and vascular invasion. In addition, Snai-1 nuclear immunoexpression was significantly related to E-cadherin loss. In conclusion, loss of epithelial proteins and/or the acquisition of mesenchymal proteins are associated with EMT and may have an important role in the evaluation of CMC patients. The unique immunoexpression pattern of Snai-1 could help to distinguish between an adenoma and a non-metastatic carcinoma and seems to be related to conversion of myoepithelial cells to a complete mesenchymal-like phenotype. Loss of E-cadherin and cytokeratin and change of immunoexpression pattern of Snai-1, N-cadherin, S100A4 and MMP-2 indicate the occurrence of EMT in canine mammary carcinomas and should result in an en bloc resection or a close follow-up.
Cite this paper
Salgado, B. , Rocha, R. , Soares, F. , Gärtner, F. , Rocha, N. (2014) Snai-1 and Epithelial-Mesenchymal Transition-Related Protein Immunoexpression in Canine Mammary Carcinomas. Advances in Breast Cancer Research, 3, 111-117. doi: 10.4236/abcr.2014.34017.
Salgado, B. , Rocha, R. , Soares, F. , Gärtner, F. , Rocha, N. (2014) Snai-1 and Epithelial-Mesenchymal Transition-Related Protein Immunoexpression in Canine Mammary Carcinomas. Advances in Breast Cancer Research, 3, 111-117. doi: 10.4236/abcr.2014.34017.
References
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http://dx.doi.org/10.1016/0021-9975(89)90022-4 [2] Larue, L. and Bellacosa, A. (2005) Epithelial-Mesenchymal Transition in Development and Cancer: Role of Phosphatidylinositol 3’ Kinase/AKT Pathways. Oncogene, 24, 7443-7454.
http://dx.doi.org/10.1038/sj.onc.1209091 [3] Jechlinger, M., Grunert, S., Tamir, I.H., Janda, E., Lüdermann, S., Waerner, T., Seither, P., Weith, A., Beug, H. and Kraut, N. (2003) Expression Profiling of Epithelial Plasticity in Tumor Progression. Oncogene, 22, 7155-7169. http://dx.doi.org/10.1038/sj.onc.1206887 [4] Thiery, J.P. (2002) Epithelial-Mesenchymal Transitions in Tumour Progression. Nature Reviews Cancer, 2, 442-454. http://dx.doi.org/10.1038/nrc822 [5] Gotzmann, J., Mikula, M., Eger, A., Schulte-Hermann, R., Foisner, R., Beug, H. and Mikulitis, W. (2004) Molecular Aspects of Epithelial Cell Plasticity: Implications for Local Tumor Invasion and Metastasis. Mutation Research, 566, 9-20. http://dx.doi.org/10.1016/S1383-5742(03)00033-4 [6] Misdorp, W., Else, R.W., Hellmen, E. and Lipscomb, T.P. (1999) Histological Classification of Mammary Tumors of the Dog and Cat. Armed Forces Institute of Pathology, Washington. [7] Elston, C.W. and Ellis, I.O. (1998) Assessment of Histological Grade. In: Systemic Pathology, 3rd Edition, Churchill Livingstone, London, 365-384. [8] Bongiovanni, L., D’Andrea, A., Romanucci, M., Malatesta, D., Candolini, M., Salda, L.D., Mechelli, L., Sforna, M. and Brachelente, C. (2013) Epithelial-Mesenchymal Transition: Immunohistochemical Investigation of Related Molecules in Canine Cutaneous Epithelial Tumours. Veterinary Dermatology, 24, 195-203. http://dx.doi.org/10.1111/j.1365-3164.2012.01116.x [9] Im, K.S., Kim, J.H., Kim, N.H., Yu, C.H., Hur, T.Y. and Sur, J.H. (2012) Possible Role of Snail Expression as a Prognostic Factor in Canine Mammary Neoplasia. Journal of Comparative Pathology, 147, 121-128. http://dx.doi.org/10.1016/j.jcpa.2011.12.002 [10] Willipinski-Stapelfeldt, B., Riethdorf, S., Assmann, V., Woefle, U., Rau, T., Sauter, G., Heukeshoven, J. and Pantel, K. (2005) Changes in Cytoskeletal Protein Composition of an Epithlelial-Mesenchymal Transition in Human Micrometastatic and Primary Breast Carcinoma Cells. Clinical Cancer Research, 11, 8006-8014. http://dx.doi.org/10.1158/1078-0432.CCR-05-0632 [11] Karantza, V. (2011) Keratins in Health and Cancer: More than Mere Epithelial Cell Markers. Oncogene, 30, 127-138. http://dx.doi.org/10.1038/onc.2010.456 [12] Rudland, S.S., Martin, L., Roshanlall, C., Winstanley, J., Leinster, S., Platt-Higgins, A., Carroll, J., West, C., Barraclough, R. and Rudland, P. (2006) Association of S100A4 and Osteopontin with Specific Prognostic Factors and Survival of Patients with Minimally Invasive Breast Cancer. Clinical Cancer Research, 12, 1192-1200. http://dx.doi.org/10.1158/1078-0432.CCR-05-1580 [13] Ismail, N.I., Kaur, G., Hashim, H. and Hassan, M.S. (2008) S100A4 Overexpression Proves to Be Independent Marker for Breast Cancer Progression. Cancer Cell International, 8, 12.
http://dx.doi.org/10.1186/1475-2867-8-12 [14] Brunetti, B., Sarli, G., Preziosi, R., Monari, I. and Benazzi, C. (2005) E-cadherin and β-Catenin Reduction Influence Invasion But Not Proliferation and Survival in Canine Malignant Mammary Tumors. Veterinary Pathology, 42, 781-787. http://dx.doi.org/10.1354/vp.42-6-781 [15] Nassar, A., Sookhan, N., Santisteban, M., Bryant, S.C., Boughey, J.C., Giorgadze, T. and Degnim, A. (2010) Diagnostic Utility of Snail in Metaplastic Breast Carcinoma. Diagnostic Pathology, 5, 76.
http://dx.doi.org/10.1186/1746-1596-5-76 [16] Gwin, K., Buell-Gutbrod, R., Tretiakova, M. and Montag, A. (2010) Epithelial-to-Mesenchymal Transition in Metaplastic Breast Carcinomas with Chondroid Differentiation: Expression of the E-Cadherin Repressor Snail. Applied Immunohistochemistry and Molecular Pathology, 19, 526-531.
http://dx.doi.org/10.1097/PAI.0b013e3181e8d54b [17] Stremmer, V., de Crane, B., Berx, G. and Behrens, J. (2008) Snail Promotes Wnt Target Gene Expression and Interacts with β-Catenin. Oncogene, 27, 5075-5080.
http://dx.doi.org/10.1038/onc.2008.140
[1] Nerurkar, V.R., Chitale, A.R., Jalnakurpar, B.V., Naik, S. and Lalitha, V.S. (1989) Comparative Pathology of Canine Mammary Turmours. Journal of Comparative Pathology, 101, 389-397.
http://dx.doi.org/10.1016/0021-9975(89)90022-4 [2] Larue, L. and Bellacosa, A. (2005) Epithelial-Mesenchymal Transition in Development and Cancer: Role of Phosphatidylinositol 3’ Kinase/AKT Pathways. Oncogene, 24, 7443-7454.
http://dx.doi.org/10.1038/sj.onc.1209091 [3] Jechlinger, M., Grunert, S., Tamir, I.H., Janda, E., Lüdermann, S., Waerner, T., Seither, P., Weith, A., Beug, H. and Kraut, N. (2003) Expression Profiling of Epithelial Plasticity in Tumor Progression. Oncogene, 22, 7155-7169. http://dx.doi.org/10.1038/sj.onc.1206887 [4] Thiery, J.P. (2002) Epithelial-Mesenchymal Transitions in Tumour Progression. Nature Reviews Cancer, 2, 442-454. http://dx.doi.org/10.1038/nrc822 [5] Gotzmann, J., Mikula, M., Eger, A., Schulte-Hermann, R., Foisner, R., Beug, H. and Mikulitis, W. (2004) Molecular Aspects of Epithelial Cell Plasticity: Implications for Local Tumor Invasion and Metastasis. Mutation Research, 566, 9-20. http://dx.doi.org/10.1016/S1383-5742(03)00033-4 [6] Misdorp, W., Else, R.W., Hellmen, E. and Lipscomb, T.P. (1999) Histological Classification of Mammary Tumors of the Dog and Cat. Armed Forces Institute of Pathology, Washington. [7] Elston, C.W. and Ellis, I.O. (1998) Assessment of Histological Grade. In: Systemic Pathology, 3rd Edition, Churchill Livingstone, London, 365-384. [8] Bongiovanni, L., D’Andrea, A., Romanucci, M., Malatesta, D., Candolini, M., Salda, L.D., Mechelli, L., Sforna, M. and Brachelente, C. (2013) Epithelial-Mesenchymal Transition: Immunohistochemical Investigation of Related Molecules in Canine Cutaneous Epithelial Tumours. Veterinary Dermatology, 24, 195-203. http://dx.doi.org/10.1111/j.1365-3164.2012.01116.x [9] Im, K.S., Kim, J.H., Kim, N.H., Yu, C.H., Hur, T.Y. and Sur, J.H. (2012) Possible Role of Snail Expression as a Prognostic Factor in Canine Mammary Neoplasia. Journal of Comparative Pathology, 147, 121-128. http://dx.doi.org/10.1016/j.jcpa.2011.12.002 [10] Willipinski-Stapelfeldt, B., Riethdorf, S., Assmann, V., Woefle, U., Rau, T., Sauter, G., Heukeshoven, J. and Pantel, K. (2005) Changes in Cytoskeletal Protein Composition of an Epithlelial-Mesenchymal Transition in Human Micrometastatic and Primary Breast Carcinoma Cells. Clinical Cancer Research, 11, 8006-8014. http://dx.doi.org/10.1158/1078-0432.CCR-05-0632 [11] Karantza, V. (2011) Keratins in Health and Cancer: More than Mere Epithelial Cell Markers. Oncogene, 30, 127-138. http://dx.doi.org/10.1038/onc.2010.456 [12] Rudland, S.S., Martin, L., Roshanlall, C., Winstanley, J., Leinster, S., Platt-Higgins, A., Carroll, J., West, C., Barraclough, R. and Rudland, P. (2006) Association of S100A4 and Osteopontin with Specific Prognostic Factors and Survival of Patients with Minimally Invasive Breast Cancer. Clinical Cancer Research, 12, 1192-1200. http://dx.doi.org/10.1158/1078-0432.CCR-05-1580 [13] Ismail, N.I., Kaur, G., Hashim, H. and Hassan, M.S. (2008) S100A4 Overexpression Proves to Be Independent Marker for Breast Cancer Progression. Cancer Cell International, 8, 12.
http://dx.doi.org/10.1186/1475-2867-8-12 [14] Brunetti, B., Sarli, G., Preziosi, R., Monari, I. and Benazzi, C. (2005) E-cadherin and β-Catenin Reduction Influence Invasion But Not Proliferation and Survival in Canine Malignant Mammary Tumors. Veterinary Pathology, 42, 781-787. http://dx.doi.org/10.1354/vp.42-6-781 [15] Nassar, A., Sookhan, N., Santisteban, M., Bryant, S.C., Boughey, J.C., Giorgadze, T. and Degnim, A. (2010) Diagnostic Utility of Snail in Metaplastic Breast Carcinoma. Diagnostic Pathology, 5, 76.
http://dx.doi.org/10.1186/1746-1596-5-76 [16] Gwin, K., Buell-Gutbrod, R., Tretiakova, M. and Montag, A. (2010) Epithelial-to-Mesenchymal Transition in Metaplastic Breast Carcinomas with Chondroid Differentiation: Expression of the E-Cadherin Repressor Snail. Applied Immunohistochemistry and Molecular Pathology, 19, 526-531.
http://dx.doi.org/10.1097/PAI.0b013e3181e8d54b [17] Stremmer, V., de Crane, B., Berx, G. and Behrens, J. (2008) Snail Promotes Wnt Target Gene Expression and Interacts with β-Catenin. Oncogene, 27, 5075-5080.
http://dx.doi.org/10.1038/onc.2008.140