The proliferative activity of 32 primary transitional cell tumours of the bladder (11G1, 10G2, 11G3; 18pTa, 9pT1, 5pT2-4) has been investigated on formalin-fixed, paraffin-embedded biopsy specimens utilizing AgNOR silver staining, PCNA immunostaining (anti PC10 monoclonal antibody) and DNA flow cytometry. The AgNOR count was highly associated with the tumour histologic grade (6.14 for G1, 7.57 for G2, 13.25 for G3; p<0.0001) or stage (6.54 for pTa, 11.33 for pT1, 13.87 for pT2-4; p<0.0001). The PCNA scores were associated with tumour histologic grade (13.62% for G1, 21.98% for G2, 37.47% for G3) and significantly different between pTa (16.22%) and pT1 (33.63%; p<0.0001) or pT2-4 (37.42%; p<0.0001) tumours, but not between pT1 and pT2-4 cases (p=0.27). Aneuploidy was detected in90% G3, 30% G2 and 54.5% G1 tumours (p<0.016); and in 80% pT2-4, 77% pT1 and 44% pTa tumours (p=0.14). A linear relationship was found between AgNOR count and PCNA scores (r=0.85, p<0.0001). The AgNOR count was significantly higher in the 19 aneuploid (10.07) then in the 13 diploid (7.51) cases (p=0.03), while PCNA scores were not significantly correlated with ploidy (20.97% for diploid vs 26.79% for aneuploid tumours; p=0.19). In the diploid tumours no relationship was found between S-phase and AgNOR count (r=0.41) or PCNA scores (r=0.18); and between Proliferative Index and AgNOR count (r=0.1) or PCNA scores (r=0.15). Our results indicate that AgNOR analysis, PCNA immunohistochemisty and DNA flow cytometry are all reliable methods for evaluating the proliferative activity of transitional cell carcinoma of the bladder and may offer useful prognostic parameters. AgNOR and PCNA in particular, have the advantage of speed and simplicity over DNA flow cytometry; they can also exclude areas of extensive necrosis and stromal or inflammatory cells and allow the simultaneous evaluation of the tumour morphology and kinetics even on very small biopsies performed.
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