EpCAM Expression in Canine Malignant Mammary Tumors - Immunohistochemical Study
DOI:
https://doi.org/10.22456/1679-9216.125007Abstract
Background: Epithelial cell adhesion molecule (EpCAM) is a glycoprotein responsible for multiple cellular functions that includes cell-to-cell adhesion and signal transduction. Studies in human breast cancer demonstrated that its overexpression has been linked to a more aggressive histological phenotype. However, in canine mammary tumors, its use is still scarce and its function, remains unknown. The main goal of the present study was to characterize the expression of EpCAM in canine mammary malignant tumors and associate its expression with clinicopathological features.
Materials, Methods & Results: Forty-eight canine mammary samples were analyzed in the present study and went through the Hematoxylin-Eosin (HE) routine technique for histopathological and immunohistochemical analysis. EpCAM expression was evaluated considering the total immunolabelling (TI) which consists of the addition of 2 parameters: proportion score (PS) and intensity score (IS). Thirteen (27.1%) cases showed weak expression, 9 (18.7%) were characterized by a moderate expression and 27 (54.2%) were classified as intense. EpCAM overexpression was described in 36 (72.9%) primary tumors and in 5 lymph node metastases (71.4%). Immunoreactivity was, mainly, characterized by a staining in the lateral cell membrane of luminal epithelial cells, both in primary tumors and lymph node metastases. The statistical analysis was performed using the SPSS Statistics, version 24. Clinicopathological features, such as histological grade (P = 0.027), intravascular emboli (P = 0.004) and lymph node metastases (P = 0.016) demonstrated to be associated with a more aggressive histological type. Intravascular emboli and lymph node metastases were also linked to a higher histological grade.
Discussion: EpCAM overexpression has been described in the literature both in canine mammary tumors and breast cancer. While the results in breast cancer could be ambiguous, depending on the tumor subtype, in the present study our findings demonstrated that overexpression was present in more than 50% of the total cases evaluated. In addition, immunoreactivity was also in accordance with the literature that describes membranous staining as the most prevalent pattern type. It was not possible to correlate overexpression with histological type and histological grade. However, those clinicopathological features are essential to evaluate tumor prognosis. For instance, our results demonstrated that histological type was statistically significant with histological grade (P = 0.027), intravascular emboli (P = 0.004) and lymph node metastases (P = 0.016). Additionally, the Nottingham system was statistically significant with intravascular emboli (P = 0.033) and lymph node metastases (P = 0.006). These findings have also been described in similar studies using canine mammary tissues in bitches. When it comes to pattern of expression in primary tumors and lymph nodes metastases, our findings described that neoplastic cell presented the same phenotype in these 2 different locations. In addition, loss of expression was also described in cluster cells within the subcapsular are which reflects the idea that the expression of EpCAM molecule changes according to time, which could be translated into the multistep process of metastases. Finally, it demonstrates that the development of neoplasia is not something static, but rather dynamic.
Keywords: EpCAM, canine, bitches, mammary tumors, neoplasia, adhesion cell molecule, immunohistochemistry.
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Araújo M.R., Campos L.C., Ferreira E. & Cassali G.D. 2015. Quantitation of the Regional Lymph Node Metastatic Burden and Prognosis in Malignant Mammary Tumors of Dogs. Journal Veterinary Intern Medicine. 29: 1360-1367. DOI: https://doi.org/10.1111/jvim.13576
Avallone G., Rasotto R., Chambers J.K., Miller A.D., Behling-Kelly E., Monti P., Berlato D., Valenti P. & Roccabianca P. 2021. Review of histologial grading systems in veterinary medicine. Veterinary Pathology. 58(5): 809-828. DOI: https://doi.org/10.1177/0300985821999831
Canadas A., Santos M., Medeiros R. & Dias-Pereira P. 2019. Influence of E-cadherin genetic variation in canine mammary tumour risk, clinicopathological features and prognosis. Veterinary Comparative Oncology. 17: 489-496. DOI: https://doi.org/10.1111/vco.12510
Coleto A.F., Wilson T.M., Soares N.P., Gundim L.F., Castro I.P., Guimarães E.C., Bandarra M.B. & Medeiros-Ronchi A.A. 2018. Prognostic Value of Occult Isolated Tumour Cells within Regional Lymph Nodes of Dogs with Malignant Mammary Tumours. Journal Comparative Pathology. 158: 32-38. DOI: https://doi.org/10.1016/j.jcpa.2017.11.001
Collivignarelli F., Tamburro R., Aste G., Falerno I., Del Signore F., Simeoni F., Patsikas M., Gianfelici J., Terragni R., Attorri V., Carluccio A. & Vignoli. M. 2021. Lymphatic Drainage Mapping with Indirect Lymphography for Canine Mammary Tumors. Animals (Basel). 11(4): 1115. DOI: https://doi.org/10.3390/ani11041115
Figuera A.C., Gomes C., Oliveira J.T., Vilhena H., Carvalheira J., Matos A.J., Pereira P.D. & Gartner F. 2014. Aberrant P-cadherin expression is associated to aggressive feline mammary carcinomas. BMC Veterinary Research. 10: 270. DOI: https://doi.org/10.1186/s12917-014-0270-z
Figueira A.C., Gomes C., Mendes N., Amorim I., Matos A.J.F., Dias-Pereira P. & Gartner F. 2016. An in vitro and in vivo characterization of the cadherin-catenin adhesion complex in a feline mammary carcinoma cell line. Clinical and Diagnostic Pathology. 1(1): 1-8. DOI: https://doi.org/10.15761/NRD.1000103
Gama A. & Schimitt F. 2012. Cadherin cell adhesion system in canine mammary cancer: a review. Veterinary Medicine International. 2012: 357187. DOI: https://doi.org/10.1155/2012/357187
Gama A., Paredes J., Gartner F., Alves A. & Schimitt F. 2008. Expression of E-cadherin, P-cadherin and beta-catenin in canine malignant mammary tumours in relation to clinicopathological parameters, proliferation and survival. Veterinary Journal. 177: 45-53. DOI: https://doi.org/10.1016/j.tvjl.2007.05.024
Gao S., Sun Y., Liu X., Zhang D & Yang X. 2017. EpCAM and COX2 expression are positively correlated in human breast cancer. Molecular Medicine Reports. 15: 3755-3760. DOI: https://doi.org/10.3892/mmr.2017.6447
Gastl G., Spizzo G., Obrist P., Dunser M. & Mikuz G. 2000. Ep-CAM overexpression in breast cancer as a predictor of survival. The Lancet. 356: 1981-1982. DOI: https://doi.org/10.1016/S0140-6736(00)03312-2
Lee S.J. & Yun C.C. 2010. Colorectal cancer cells - Proliferation, survival and invasion by lysophosphatidic acid. The International Journal of Biochemistry & Cell Biology. 42: 1907-1910. DOI: https://doi.org/10.1016/j.biocel.2010.09.021
Matos A.J.F., Faustino A.M.R., Lopes C., Rutteman R. & Gärtner F. 2006. Detection of lymph node micrometastases in malignant tumours in dogs by cytokeratin immunostaining. Veterinary Record. 158: 626-630. DOI: https://doi.org/10.1136/vr.158.18.626
Michishita M. 2020. Understanding of tumourigenesis in canine mammary tumours based on cancer stem cell research. Veterinary Journal. 265: 105560. DOI: https://doi.org/10.1016/j.tvjl.2020.105560
Misdorp W. 1999. Histological Classification of Mammary Tumors od the Dog and the Cat. Word Health Organization International Histological Classification of Tumors of Domestic Animals Second Series. 7: 1-59.
Peña L., Gama A., Goldschmidt M.H., Abadie J., Benazzi C., Castagnaro M., Diez L., Gartner F., Hellmen E., Kiupel M., Millan Y., Miller M.A., Nguyen F., Poli A., Sarli G., Zappulli V. & De Las Mulas J.M. 2014. Canine mammary tumors: a review and consensus of standard guidelines on epithelial and myoepithelial phenotype markers, HER2, and hormone receptor assessment using immunohistochemistry. Veterinary Pathology. 51: 127-145. DOI: https://doi.org/10.1177/0300985813509388
Pereira P.D. & Gärtner F. 2003. Expression of E-cadherin in normal, hyperplastic and neoplastic feline mammary tissue. Veterinary Record. 153: 297-302. DOI: https://doi.org/10.1136/vr.153.10.297
Redig A.J. & Mcallister S.S. 2013. Breast cancer as a systemic disease: a view of metastasis. Journal Intern Medicine. 274: 113-126. DOI: https://doi.org/10.1111/joim.12084
Rocha A.A., Carvalheira J. & Gärtner F. 2020. Alpha-catenin, beta-catenin and P-120-catenin immunoexpression in canine mammary tissues and their relationship with E-cadherin. Research Veterinary Science. 130: 197-202. DOI: https://doi.org/10.1016/j.rvsc.2020.03.002
Sarli G., Preziosi R., De Tolla L., Brunetti B. & Benazzi C. 2004. E-cadherin immunoreactivity in canine mammary tumors. Journal of Veterinary Diagnostic Investigation. 16: 542-547. DOI: https://doi.org/10.1177/104063870401600608
Sarrio D., Perez-Mies B., Hardisson D., Moreno-Bueno G., Suarez A., Cano A., Martin-Perez J., Gamallo C. & Palacios J. 2004. Cytoplasmic localization of p120ctn and E-cadherin loss characterize lobular breast carcinoma from preinvasive to metastatic lesions. Oncogene. 23: 3272-3283. DOI: https://doi.org/10.1038/sj.onc.1207439
Schnell U., Cirulli V. & Giepmans B.N. 2013. EpCAM: structure and function in health and disease. Biochimica et Biophysica Acta (BBA) – Biomembranes. 1828: 1989-2001. DOI: https://doi.org/10.1016/j.bbamem.2013.04.018
Soysal S.D., Muenst S., Barbie T., Fleming T., Gao F., Spizzo G., Oertli D., Viehl C.T., Obermann E.C. & Gillanders W.E. 2013. EpCAM expression varies significantly and is differentially associated with prognosis in the luminal B HER2(+), basal-like, and HER2 intrinsic subtypes of breast cancer. British Journal of Cancer. 108: 1480-1487. DOI: https://doi.org/10.1038/bjc.2013.80
Spizzo G., Fong D., Wurm M., Ensinger C., Obrist P., Hofer C., Mazzoleni G., Gastl G. & Went P. 2011. EpCAM expression in primary tumour tissues and metastases: an immunohistochemical analysis. Journal of Clinical Pathology. 64: 415-420. DOI: https://doi.org/10.1136/jcp.2011.090274
Szczubial M. & Lopuszynski W. 2011. Prognostic value of regional lymph node status in canine mammary carcinomas. Veterinary Comparative Oncology. 9: 296-303. DOI: https://doi.org/10.1111/j.1476-5829.2011.00268.x
Thamm D.H., Hayes D.F., Meuten T., Laver T. & Thomas D.G. 2016. Epithelial Cell Adhesion Molecule Expression in Canine Tumours. Journal Comparative Pathology. 155: 299-304. DOI: https://doi.org/10.1016/j.jcpa.2016.07.010
Valdivia G., Alonso-Diez A., Perez-Alenza D. & Peña L. 2021. From Conventional to Precision Therapy in Canine Mammary Cancer: A Comprehensive Review. Frontiers in Veterinary Science. 8: 623800. DOI: https://doi.org/10.3389/fvets.2021.623800
Van Der Gun B.T, Melchers L.J., Ruiters M.H., De Leij L.F, Mclaughlin P.M. & Rots M.G. 2010. EpCAM in carcinogenesis: the good, the bad or the ugly. Carcinogenesis. 31: 1913-21. DOI: https://doi.org/10.1093/carcin/bgq187
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