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Home » Archive » 2008

TDK conference 2008

Immunohistochemical examination of normal and tumourous mammary glands in canine
Szabára Ágnes - year 5
Department of Pathology and Forensic Veterinary Medicine
Supervisor: Dr. Csaba Jakab

Abstract:

The immunohistochemistry is an indispensable examining method in the tumour pathology. Histogenesis of the primer and seconder tumour and the characteristic antigen profil of the tumours are specifiable with that. Molecular background of the tumours and the tumour invasion are qualifiable. We use several immunohistochemical markers adjudication of prognosis of the mammary tumours with the help of which biological aggressiveness and the malignant potential of the tumours are specifiable.Biopsy samples (N = 166) of canine normal mammary gland (n = 15) and mammary gland lesions were submitted for diagnosis to the Szent István University, Faculty of Veterinary Science, Department of Pathology and Forensic Veterinary Medicine. Lesions were classified by the World Health Organisation system as lobular hyperplasia (n = 15), simple adenoma (n = 15), complex adenoma (n = 15), benign mixed tumour (n = 15), non-infiltrating carcinoma in situ (n = 15), complex carcinoma without metastasis (n = 15), complex carcinoma with metastasis (n = 5), grade I tubulopapillary infiltrating simple carcinoma (n = 15), grade III solid invasive simple carcinoma with lymphogen invasion (n = 15), reccurence simple carcinoma (n = 5), locoregional micrometastases of simple carcinomas (n = 8), and distant (lungs) metastases of the solid infiltrating simple carcinoma (n = 10). Serial sections (3 – 4 μm) were initially dewaxed in xylene and graded ethanol. After treatment with appropriate antigen retrieval, the sections were incubated with primary human antibodies against CKAE1-AE3, CK14, CK5/6, alpha-SMA, p63, calponin, vimentin, S-100 protein, p53, Ki-67, PCNA, E-cadherin, HER-2, claudin-5, estrogen and progesterone receptors for 60 min at room temperature. Immunohistochemical labelling was performed using the streptavidin-peroxidase procedure. The chromogen substrate was 3, 3’-diaminobenzidine tetrahydrochloride (DAB substrate-chromogen). Sections were counterstained with Mayer’s haematoxylin. In the first part of the study we inestigated the immunohistochemical expression pattern of the canine normal mammary gland with antibodies which present cross reactivity with human and animal antigensand we found internal positive controls. These positive controls helped the immunhistochemical investigation of the canine mammary gland tumours. In the second part of the immunohistochemical research we analised the relationship between the prognosis of the canine mammary gland tumours and the expression of the several markers, and the histological type, grade, nodal-state and reccurence of these tumours.



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