The levels of 26 kDa-soluble (S) and 30 kDa-membrane-bound (MB) catechol-O-methyltransferase (COMT) polypeptides were determined in paired samples from normal and neoplastic breast tissue of 32 patients with breast cancer. Immunohistochemical staining showed that the COMT reaction in normal mammary tissue was restricted to the epithelial cells in the ducti and lobuli, whereas in the tumors a strong reaction was also seen in the malignant cells. The amounts of COMT proteins in tumors could not be correlated with various clinical or pathological parameters. Quantitative immunoblotting analysis revealed that the total amount of COMT proteins in tumors was more than 50% higher than in respective normal samples in 26 out of 32 patients. Five cases showed less than a 50% difference and in one case less COMT was detected in the tumor. In most cases the amount of both S- and MB-COMT forms was increased. The average amount of total COMT was 178 +/- 57 pg/microg total protein in normal tissue and 566 +/- 94 pg/microg total protein in tumor. Respective values for S-COMT were 137 +/- 52 pg/microg total protein in normal tissue and 369 +/- 62 pg/microg total protein in tumor and for MB-COMT 41 +/- 10 and 197 +/- 41 pg/microg total protein, respectively. Analysis of COMT-specific transcripts suggested that the COMT enzyme level in tumors is determined in some cases by transcriptional and in some cases by post-transcriptional mechanisms.