BACKGROUND Transitional cell carcinoma (TCC) of the bladder displays an unpredictable biologic behavior and the morphologic methods of grading tumor malignancy are often insufficient to predict the clinical outcome of patients with TCC of the bladder. Thus, the new indicator should reliably reflect prognosis. In this study, the authors determined the prognostic significance of proliferating cell nuclear antigen (PCNA), p53 protein, and nm23 protein, as well as nuclear DNA content in specimens with TCC of the bladder. METHODS Paraffin embedded materials taken from 77 patients with nonmetastatic untreated TCC of the bladder (classified as pTa-3b, NO, MO) treated with total cystectomy were employed in this study. PCNA expression, p53 protein and nm23 protein immunoreactivities, and the parameters for nuclear DNA content such as 2c deviation index (2cDI) and 5c exceeding rate (5cER) were evaluated using a computer-assisted image analyzer, and the results were compared with histologic findings and clinical outcome. RESULTS PCNA expression positively correlated with p53 protein and nm23 protein immunoreactivities, 2cDI value, and 5cER. In addition, histologic grade positively correlated with all of these five parameters. Similarly, pT category and disease progression positively correlated with all of the five parameters, except for nm23 protein immunoreactivity. In tumors with high genetic instability as judged by 2cDI and 5cER, both PCNA expression and p53 protein immunoreactivity were elevated, whereas nm23 protein immunoreactivity was not. However, the percent coefficient of variation in PCNA expression was smaller than that observed in p53 protein immunoreactivity in each group of genetic instabilities. In univariate analysis, prognostic potential was found with histologic grade, pT category, PCNA expression, p53 protein immunoreactivity, 2cDI value, and 5cER, but was not noted in nm23 protein immunoreactivity. Multivariate analysis indicated that quantity and intensity of PCNA expression (chi 2 = 8; P = 0.0047 for quantity and chi 2 = 8.71; P = 0.0032 for intensity) and 2cDI value (chi 2 = 5.52; P = 0.0019) were independent variables of histologic grade and pT category when predicting survival. However, p53 protein and nm23 protein immunoreactivities and 5cER were not of independent significance. CONCLUSIONS The tumor growth fraction as assessed by PCNA immunostaining is an independently significant predictor for survival of patients with TCC of the bladder.