Quantification of DNA and AgNORs correlating to cyclin D1-immunoexpression in human breast cancer

Abstract

Background: Assessment of cell proliferation through the evaluation of cyclin D1, AgNORs, and DNA content provides a promising approach to the prognosis of breast cancer. Therefore, this work aimed to evaluate the prognostic significance of AgNORs and DNA image cytometry, and their relation to cyclin D1 immunoreactivity in breast cancer. Subjects and methods: This study was carried out on 50 prospective biopsies taken from patients with breast tumors. They were divided into control (10 cases) and benign (10 cases) as well as malignant (30 cases) groups. Computer-assisted image analysis cytometry is applied to evaluate the count of positive reactions of AgNORs and cyclin D1, and an integrated optical density (IOD) technique is used to quantify DNA content in the tissues stained by Feulgen stain and to assess the density of cyclin D1 immunoreactivity. Results: The comparison between the mean count of cyclin D1 positive reaction (Mean ± SD: 31.10 ± 26.39) and AgNORs dots (Mean ± SD: 4.64 ± 2.77) revealed a statistically significant difference (P <0.001) in the normal group and in the malignant group as well as in total cases. The association between the IOD parameter for Feulgen stain (Mean ± SD: 77.35 ± 27.99) and cyclin D1 immunostaining (Mean ± SD: 151.90 ± 37.38) showed a statistically significant difference throughout all of the individual groups (P <0.001). Conclusions: These results indicate that image cytometry may be a valuable method for quantitating DNA, AgNORs, and cyclin D1-immunostaining, and that provides valuable information that may have prognostic significance for patients with breast cancer.