The cytosolic phospholipase A2 (PLA2) was characterized in the human epithelial carcinoma cell line HEp-2 by its apparent molecular mass (about 80 kDa); its in vitro activation by micromolar concentrations of calcium; and its calcium-dependent association with cellular membranes. The activity of this enzyme was induced by an overnight incubation with tumor necrosis factor-alpha (TNF-alpha). Glucocorticoids only moderately reduced PLA2 activity in control cells, but completely inhibited the TNF-alpha-induced increase in the activity of the high-molecular-weight cytosolic PLA2.