Silver-Stained Nucleolar Organizer Regions in Normal and Dysplastic Cervical Lesions: Correlation with DNA Ploidy and S-Phase Fraction by Flow Cytometry

Abstract

Objectives: To investigate the prognostic significance of DNA ploidy and silver-stained nucleolar organizer regions (AgNOR), their relation to the cytological grades of cervical dysplasia and correlation of cells in the S-phase of the cell cycle and DNA ploidy with AgNOR count. Methods: Multiparametric DNA flow-cytometric analysis and AgNOR count were performed on cervical smear samples, cytologically diagnosed as normal (n = 196), atypical squamous cells of unknown significance (ASCUS, n = 98) and various grades of cervical dysplasia (n = 127). Results: Among the cytological grade, aneuploid cases from low-grade squamous intraepithelial lesion (mild) and high-grade squamous intraepithelial lesion (moderate and severe) groups were showing high AgNOR counts (ranges 4.33–5.86 and 5.91–7.42, respectively). Aneuploid cases from the ASCUS group also presented high AgNOR (range 3.72–4.93). The high percentage of cells in the S-phase (>12%) correlated with these 2 parameters in the ASCUS and dysplasia groups. After grouping the cases into those with high (>4.0) and low (<4.0) AgNOR counts, significant correlations in the group with high AgNOR count with DNA ploidy, S-phase and cytological grade were recorded (p < 0.001). Results obtained from follow-up study revealed that cases with aneuploidy, high AgNOR count and S-phase fraction presented with a progressive trend of disease. Conclusions: Thus, the study shows that high AgNOR count, S-phase fraction and aneuploid DNA have prognostic significance in the early detection of cervical cancer.

AgNOR polymorphism association with squamous intraepithelial lesions and invasive carcinoma with HPV infection

“…Atypical polymorphism showed a significant increase in patients with LSIL, HSIL and SCC. This increase may be explained by the relationship between morphological changes in the AgNORs and high transcriptional activity due to proliferative potential, increased activity of rRNA genes, ploidy and aneuploid DNA 15,39 or infection of the cells with HPV. 40 In this work, all samples were infected with high-risk HPV types.…”

“…[11][12][13] It has also been shown that high AgNOR count, 14 S-phase fraction and aneuploid DNA have prognostic significance in the early detection of cervical cancer. 15 However, at present it has also been described that the increase in polymorphism of AgNORs in cervical biopsies could be used as a possible marker for histopathological diagnosis. 16,17 Furthermore, only a few reports have been published concerning the variability of AgNORs in cervical cytology in women with cervical neoplastic damage, suggesting that polymorphism of AgNORs may be related to neoplastic development.…”

T lymphocyte Subsets Determination and DNA Ploidy Analysis in the Differential Diagnosis between Benign and Malignant Ascites

“…Usually 10,000 nuclei were counted and DNA ploidy was expressedby the DI. The DI was calculated according to the principles recommended by consensus (5). DI representsthe ratio of the aneuploid G 1 -G 0 -DNA peak channel to the diploid G 1 -G 0 -DNA peak channel.…”

Nucleus Detection in Cervical Samples Stained With AgNOR

“…Results demonstrated a progressive increase in AgNOR counts when the severity of the lesions increased, mainly in the differentiation between low and high-grade SIL. In addition, on [20] and [21] they analyzed the AgNOR count as a marker of malignancy progression. As the count shows the intensity of cell proliferation, this method can be used in differentiating between low and high degree of cervical dysplasia, as well as between lesions with high and low potential for malignant transformation.…”