CYTOTOXICITY OF LAZAROID U-75412E IN HUMAN EPITHELIAL-CELL LINE (WISH)

The 21-aminosteroids (or lazaroids) are a recently synthesized class o f compounds demonstrated to protect tissue against damage induced by t rauma and/or ischemia. Currently, very little is known about the biolo gical effects of lazaroids. In this work the action of lazaroid U-7541 2E on a human epithelial cell line (Wish) was evaluated. The data obta ined showed an inhibition of cell growth and a dose- and time-dependen t decrease of cell viability. Furthermore, a dose- and time-dependent increase of cells in the G2/M phase with the appearence of apoptotic c ells was observed by flow cytometric analysis. Nuclear fragmentation w as also evident. Lactate dehydrogenase release and scanning electron m icroscopy experiments suggested that plasma membrane integrity was alt ered by this compound. The immunofluorescence technique and transmissi on electron microscopy images also showed intracellular damage, such a s alteration of microtubular arrangement, mitochondrial swelling and t he presence of vacuoles. This study demonstrated that 1 mu M U-75412E was unable to modify these parameters, while higher concentrations (6- 75 mu M) had a cytotoxic effect on Wish cells.