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CA 15–3 cell lines and tissue expression in canine mammary cancer and the correlation between serum levels and tumour histological grade

Elisabetta Manuali1*, Antonio De Giuseppe1, Francesco Feliziani1, Katia Forti1, Cristina Casciari1, Maria Chiara Marchesi2, Eugenio Pacifico3, Karol M Pawłowski4, Kinga Majchrzak4 and Magdalena Król4

Author Affiliations

1 Laboratory of Histopathology and Electron Microscopy, Istituto Zooprofilattico Sperimentale dell’Umbria e delle Marche, via G. Salvemini, 1 – 06126, Perugia, Italy

2 Department of Veterinary Pathology, Diagnostic and Clinic, Faculty of Veterinary Medicine, University of Perugia, via S. Costanzo, 4 – 06126, Perugia, Italy

3 Hematology and Clinical Pathology Service, S. Maria della Misericordia - Perugia Hospital, S. Andrea delle Fratte, Perugia, 06156, Italy

4 Department of Physiological Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences - WULS, Nowoursynowska 159, Warsaw, 02-776, Poland

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BMC Veterinary Research 2012, 8:86  doi:10.1186/1746-6148-8-86

Published: 22 June 2012



Mammary tumours are the most common malignancy diagnosed in female dogs and a significant cause of mortality and morbidity in this species. Carbohydrate antigen (CA) 15–3 is a mucinous glycoprotein aberrantly over-expressed in human mammary neoplasms and one of the most widely used serum tumour markers in women with breast cancer. The aim of this study was to investigate the antigenic analogies of human and canine CA 15–3 and to assess its expression in canine mammary cancer tissues and cell lines. Immunohistochemical expression of CA 15–3 was evaluated in 7 canine mammary cancer cell lines and 50 malignant mammary tumours. As a positive control, the human breast carcinoma cell line MCF7 and tissue were used. To assess CA 15–3 staining, a semi-quantitative method was applied. To confirm the specificity and cross-reactivity of an anti-human CA 15–3 antibody to canine tissues, an immunoblot analysis was performed. We also investigated serum CA 15–3 activity to establish whether its expression could be assigned to several tumour characteristics to evaluate its potential use as a serum tumour marker in the canine mammary oncology field.


Immunocytochemical analysis revealed CA 15–3 expression in all examined canine mammary cancer cell lines, whereas its expression was confirmed by immunoblot only in the most invasive cells (CMT-W1, CMT-W1M, CMT-W2 and CMT-W2M). In the tissue, an immunohistochemical staining pattern was observed in 34 (68%) of the malignant tumours. A high statistical correlation (p = 0.0019) between serum CA 15–3 levels and the degree of tumour proliferation and differentiation was shown, which indicates that the values of this serum marker increase as the tumour stage progresses.


The results of this study reveal that CA 15–3 is expressed in both canine mammary tumour cell lines and tissues and that serum levels significantly correlate with the histological grade of the malignancy.