Introduction: In human breast cancer, phosphatidylinositol-3 kinase (PI3K) mutation is involved in tumour growth and resistance to hormone therapy. The aim of the present study was to standardize immunohistochemistry (IHC) by anti-PI3K antibody in canine mammary samples, and to evaluate the different immunohistochemical expression in tumour progression. Materials and Methods: To assess the anti-PI3K antibody (clone D-9) specificity a western blot (WB) analysis was performed in the salivary gland and in normal canine mammary tissue. Samples from three normal mammary glands (NMGs), five benign (B) and 13 malignant (M) tumours were available for IHC. Immunohistochemical labelling was interpreted independently by two authors and the expression was evaluated by a semiquantitative method (<50% or >50%) and according to a labelling intensity scale from 0 to 3. Results: WB analysis revealed two major bands of expected molecular weight (83 and 50 kDa) of PI3K isoforms. PI3K expression was >50% in all samples. Low (0 and 1) and high (2 and 3) labelling intensity was present respectively in 1 and 2 NMGs, 1 and 4 B and 2 and 11 M tumours. Conclusions: In this investigation, the anti-PI3K antibody (clone D-9) results were specific for canine mammary tissue. However, since immunohistochemical assessment of PI3K expression showed low variation in both extension and intensity from NMG to M tumours, IHC may not be the best technique to investigate the role of PI3K in tumour progression.

Is immunohistochemistry the best method to investigate phosphatidylinositol-3 kinase (PI3K) expression in canine mammary samples?

BEHA, GERMANA;AVALLONE, GIANCARLO;BERNARDINI, CHIARA;FORNI, MONICA;BENAZZI, CINZIA;SARLI, GIUSEPPE
2017

Abstract

Introduction: In human breast cancer, phosphatidylinositol-3 kinase (PI3K) mutation is involved in tumour growth and resistance to hormone therapy. The aim of the present study was to standardize immunohistochemistry (IHC) by anti-PI3K antibody in canine mammary samples, and to evaluate the different immunohistochemical expression in tumour progression. Materials and Methods: To assess the anti-PI3K antibody (clone D-9) specificity a western blot (WB) analysis was performed in the salivary gland and in normal canine mammary tissue. Samples from three normal mammary glands (NMGs), five benign (B) and 13 malignant (M) tumours were available for IHC. Immunohistochemical labelling was interpreted independently by two authors and the expression was evaluated by a semiquantitative method (<50% or >50%) and according to a labelling intensity scale from 0 to 3. Results: WB analysis revealed two major bands of expected molecular weight (83 and 50 kDa) of PI3K isoforms. PI3K expression was >50% in all samples. Low (0 and 1) and high (2 and 3) labelling intensity was present respectively in 1 and 2 NMGs, 1 and 4 B and 2 and 11 M tumours. Conclusions: In this investigation, the anti-PI3K antibody (clone D-9) results were specific for canine mammary tissue. However, since immunohistochemical assessment of PI3K expression showed low variation in both extension and intensity from NMG to M tumours, IHC may not be the best technique to investigate the role of PI3K in tumour progression.
JOURNAL OF COMPARATIVE PATHOLOGY
G, . BEHA; REYES CASTELLANOS, G.; Avallone, G.; Tolla, L. DE; Bernardini, C.; Forni, M.; Benazzi, C.; Sarli, G.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11585/590617
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