We used an immunohistochemical method to determine the concentrations of p53 and proliferating cell nuclear antigen (PCNA) to evaluate their usefulness as a predictor of malignancy and examined the relationship between PCNA and p53 in carcinomas of the extrahepatic bile duct (EHBD). Paraffin-embedded specimens from 46 patients were immunostained for PCNA and p53 using PC10 and DO7 monoclonal antibodies, respectively. The PCNA labeling index (LI) was closely associated with the stages of the tumor and depth of invasion (p < 0.05). The cumulative survival rate of patients with a low PCNA LI (LI < 47%) was found to be significantly better than that of patients with a high PCNA LI (LI > or = 47%) in all cases and patients with advanced cancer by univariate analysis (p < 0.05), but PCNA LI was not an independent prognostic factor in multivariate analysis. We detected p53 in 37% of the EHBD cancers. We also found that p53 positivity was not related to the percentage of PCNA-labelled cells or survival. The results suggest that PCNA immunoreactivity may be a useful predictor of malignancy in patients with EHBD carcinomas.