There are two isoforms of cyclooxygenase (COX), COX-1 and COX-2. Recent epidemiological and experimental studies indicated a close relationship between COXs and the pathogenesis of colorectal cancer. The purpose of this study was to investigate the possible roles of both isoforms in the proliferation of colon carcinoma cells. A human colon carcinoma cell line, COLO 320DM, was transfected with an eukaryotic expression vector carrying cDNA of either COX-1 or COX-2, the expression of which was driven by a powerful elongation factor-1alpha promoter in pEF-BOS. Both COX-1- and COX-2-expressing cells possessed a similar enzyme activity, 8-10 nmol/10 min per mg protein. Growth rates of both cell lines were stimulated by about 2-fold during a course of culture for 7 days as compared with mock-transfected cells. Although COX-1 and COX-2 are believed to have fundamentally different biological roles, essentially no differences in growth stimulation were observed between the COX-1 and COX-2 overexpressions in our experiments. The reason may be explained by high levels of COX expression, and subtle differences between the both cell lines would be possibly apparent by lower expression levels. The stimulated growth of the COX-transfected cells was accompanied by increased DNA synthesis as assessed by [3H]thymidine incorporation. Furthermore, expression of epidermal growth factor receptor was markedly increased in these cells as examined by reverse transcription-polymerase chain reaction. A COX inhibitor, indomethacin, suppressed the stimulated growth, increased DNA synthesis and induction of epidermal growth factor receptor in COX-1- and COX-2-transfected cells.