PCNA immunostaining as a valid alternative to tritiated thymidine-autoradiography to detect proliferative cell fraction in transitional cell bladder carcinomas

BACKGROUND: The thymidine labeling index (T-LI) evaluated with the autoradiographic technique on fresh specimens is the most objective and specific marker of tumor proliferative cell fraction. In this study we investigated T-LI and PCNA expression in 68 cases of transitional cell bladder carcinomas in order to evaluate whether PCNA immunostaining in paraffin sections could be a valid alternative to autoradiographic methods for cell kinetic detection. Immunohistochemistry was applied after different fixatives: usual formalin, neutral 4% buffered formalin and Bouin. The results revealed that only in buffered formalin-fixed tumor specimens, considering the most darkly stained cells, the PCNA-LI in all cases overlapped with T-LI. Higher values than those observed with autoradiography were detected in PCNA values obtained after usual formalin fixation whereas no or faint PCNA staining was seen in material fixed in Bouin. Thus, only cells with strong PCNA staining can be considered as significant in evaluating the proliferative activity in transitional cell bladder carcinoma. By using immunohistochemical methods it should thus be possible to overcome many of the inherent difficulties of autoradiography and likewise to obtain accurate and significant information on the biological aggressiveness of bladder tumors and consequently of patient prognosis.