Interphase argyrophilic nucleolar organiser regions and nucleolar counts in transitional cell bladder tumours

Abstract

The natural course of transitional cell bladder cancer is highly variable. The clinical approach to treatment is based mainly on the tumour stage and grade.1 2 However, the data obtained from these parameters are insuYcient to predict the biological behaviour of an individual tumour. Therefore, additional criteria have been proposed as potential instruments for more accurate grading.3 Argyrophilic nucleolar organiser regions (AgNORs) are segments of DNA that are sites of ribosomal RNA transcription. Counting silver stained interphase AgNORs provides an estimate of cellular proliferation, and has been found to correlate with tumour grade and stage, and to bear independent predictive value in patients with urinary bladder carcinoma. 4 However, the data obtained are controversial. 5-7The aim of our study was to estimate how the AgNOR and nucleolar counts in the tumour cells of transitional cell bladder carcinoma correlate with patients' sex, tumour grade, disease stage, recurrence pattern, and tumour related survival. Materials and methodsOur study comprised 62 patients (48 men and 14 women), mean age 63.9 years (range, 32-88), with primary transitional cell bladder carcinoma who had been operated upon in the urological department, State Pavlov Medical University between 1985 and 1990. Patients were included if their specimens contained sufficient tumour material for AgNOR staining and if they could be followed for at least five years. Patients with invasive neoplasia were treated by radical and partial cystectomy. In cases with superficial tumours, transurethral resection was performed.Follow up examination of patients with preserved bladder included cystoscopy every three months during the first year after surgery, cystoscopy every six months for the next two years, and then ultrasound sonography of the bladder followed by cystoscopy in suspect cases every one to two years. Thirty eight patients survived five years. Thirty seven patients developed recurrences, the mean (SD) recurrence rate being 24.8 (24.1) months (range, 2-90). TISSUEThe tumour samples were fixed in 10% buVered formalin and embedded in paraYn wax. Sections (5 µm thick) were stained with haematoxylin and eosin. The tumours were graded as recommended by Mostofi and colleagues 8 and staged postoperatively according to the TNM staging system. 9 In 17 cases, specimens were too small to confirm the tumour stage.AgNOR STAINING AND COUNTING Sections (5 µm thick) cut in series with the haematoxylin and eosin stained sections were fixed in methanol/acetic glacial acid mixture (3/1) for 15 minutes and rinsed thoroughly with distilled water. After air drying, the sections were transferred to 2% formic acid and incubated for 10 minutes, rinsed again, and impregnated with silver nitrate solution, according to Howell and Black, 10 with a slight modification, namely: two drops of gelatin solution were placed on the slide and mixed with four drops of freshly prepared 50% aqueous AgNO 3 solution. The slides were covered

Two-dimensional agnor evaluation as a prognostic variable in urinary bladder carcinoma: A different approach via total agnor area/nucleus area per cell

“…According to the data, we agreed with other researchers that the mean TAN/N per cell is not a useful tool for neither diagnostic nor prognostic purposes in TCC of urinary bladder (Iwata et al, 1995;Korneyev et al, 2000), while some previous data has not been consistent (Shiina et al, 1996;Pich et al, 2000). In contrast, by evaluating the TAA/NA ratio, and using our new software, we distinguished all groups from each other with high significance via mean TAA/NA proportion per cell.…”

“…However, these markers are limited as indicators of prognosis (Stein et al, 1998;Han and Shoenberg, 2000). Although the AgNOR amount is a direct parameter of the cells proliferative activity in many cancer types (Shiina et al, 1996), in bladder cancer the correlation between AgNOR amount (number or area) and tumor behavior remains controversial (Tomobe et al, 1999;Korneyev et al, 2000;Han and Shoenberg, 2000;Pich et al, 2000). Therefore, we have suggested that the main problem may be in the method(s) of measuring AgNOR or means of evaluating the data, which are obtained by counting or area measuring.…”

Prognostic Value of Microvessel Density in Tumor and Peritumoral Area as Evaluated by CD31 Protein Expression and Argyrophilic Nucleolar Organizer Region Count in Endothelial Cells in Uterine Leiomyosarcoma

“…AgNORs of human endotheliocytes were stained with 50% AgNO 3 solution (Bio-Optica, Milan) according to Howell and Black, 1980 [ 32 ], as modified by Korneyev et al, 2000 [ 33 ]. The main difference of this modification is that before silver staining of preparations, the latter were treated with 2N formic acid solution for 20 min to reduce deposition of silver outside nucleoli and facilitate the count of argyrophilic granules.…”

Chromatin Structure and Epigenetics of Tumour Cells: A Review

“…For example, the number of nuclei-related AgNORs (argyrophilic nucleolar organizer region) correlates with tumour grade [124]. Moreover, AgNOR scores in bladder carcinoma cells reflect variations in tumour biological behavior, although the clinical value of this technique is still limited [124,125]. However, other reports claim that AgNORs score is an excellent index for determining the tumour grade [126,127].…”