Forty-three cases of gallbladder cancer were investigated for c-erbB-2 gene amplification and c-erbB-2 protein over-expression using a combined polymerase chain reaction (PCR) and immunohistochemical approach. Thirty out of 43 cases (69.6%) demonstrated c-erbB-2 gene amplification, the positive rates being 50% and 77.4% for twelve early cancers and thirty-one advanced cancers, respectively (P < 0.05). However, there was no statistically significant correlation between c-erbB-2 gene amplification and histologic grade of differentiation or lymph node metastasis. Fourteen out of 43 cases (32.6%) showed positive immunoreactivity reflecting c-erbB-2 protein over-expression but again no statistically significant correlation was found with grade of differentiation, invasion or lymph node metastasis. Neither the c-erbB-2 gene nor the protein revealed any close relation to prognosis. In contrast, histopathologic findings for histologic grade of differentiation, invasion grade and lymph node metastasis showed good correlations to prognosis and between themselves. In conclusion, while c-erbB-2 gene and protein in gallbladder cancers might be related to invasiveness, they are not applicable as predictive factors for prognosis.