DEMONSTRATION OF SILVER-STAINED NUCLEOLAR ORGANIZER REGION ASSOCIATEDPROTEINS (AGNORS) AFTER WET AUTOCLAVE PRETREATMENT IN BREAST-CARCINOMA - CORRELATION TO TUMOR STAGE AND LONG-TERM SURVIVAL

Argyrophilic nucleolar organizer region associated proteins (AgNORs) a re known to reflect cellular and nucleolar activity. Due to a novel st aining procedure, which substantially improves visualisation of AgNORs on formalin-fixed and paraffin-embedded material, AgNORs can be relia bly demonstrated as true substructures of the nucleoli. The aim of the present study was to apply a standardized morphometric AgNOR quantifi cation on a large series of breast carcinomas with regard to its progn ostic relevance. AgNOR quantity was evaluated on archival tumor tissue s of 115 adenocarcinomas of the breast treated with the wet autoclave method prior to standardized silver-staining and morphometric analysis . AgNOR parameters were correlated to prognostic features (steroid hor monal receptor status, tumor type, tumor size, histological grading, p TNM, and UICC stage) carrying out both univariate and multivariate sur vival analyses. AgNOR number and area were proven to be statistically significantly related (Pearson correlation coefficient: 0.67, Bonferro ni adjusted P = 0.0001). Almost all AgNOR parameters, in particular CV (coefficient of variation) of corrected area (delta-area) and CV of n umber, were statistically significantly correlated to estrogen and pro gesterone receptor status as well as histological grading of tumors. I ncreased AgNOR parameters were statistically significantly associated with early tumor relapse and cancer related death. Univariate and mult ivariate analysis by means of Cox regression revealed independent prog nostic significance for CV of delta-area and number of AgNORs. Various AgNOR parameters (CV of number, CV of delta-area, CV of area, mean de lta-area, and mean area of AgNORs per nucleus) determined on wet autoc lave pre-treated formalin-fixed and paraffin-embedded breast cancer ti ssues are statistically highly significantly associated with the progn ostic outcome, independently predicting tumor-free and overall surviva l.