The effects of tumor necrosis factor-alpha (TNF-alpha) on cultured human umbilical vein endothelial cells (EC) and five cancer cell lines, A549, ME180, A2780, KURAMOCHI, and Hela, were compared. While A549, A2780, KURAMOCHI, and Hela cells were fairly resistant to the cytolytic effects of TNF-alpha, ME180 cells were sensitive. EC were also less sensitive to TNF-alpha than ME180 cells as judged by the viability of individual cells and by the release of lactate dehydrogenase (LDH) into the medium. Manganese superoxide dismutase (Mn-SOD) was markedly induced by these cytokines in EC and in A549 cells but not in ME180 cells. The levels of Mn-SOD in the conditioned medium of EC were dramatically increased after stimulation with cytokines, whereas those in ME180 and A549 cells were relatively low. The amount of Mn-SOD released appears to be comparable to that from cells lysed by other means. Immunoblot analysis of Mn-SOD in the medium showed that the molecular mass of the immunoreactive protein was the same as mitochondrial Mn-SOD, indicating that no proteolysis had occurred. These data suggest that in vivo the TNF-alpha produced by cancer cells may induce Mn-SOD in vascular endothelial cells, resulting in release of a relatively large amount of this protein into the serum.