Increase in hormonal activation of adenylate cyclase by treatment of cultured cells with N-alkyl,N-nitrosourea.

Treatment of cultured normal rat kidney cells with the nitrosourea-containing compounds streptozotocin, chlorozotocin, or 1-(2-chloroethyl)-3-(4-methylcyclohexyl)-1-nitrosourea resulted in a time-dependent potentiation in the ability of prostaglandin E1 and (-)-isoproterenol to elevate intracellular cAMP levels. This hormone response increased at 4 hours and reached a maximum at 15--25 hours after addition of the nitrosoureas. Basal cAMP levels were not affected. The greater response was apparently due to an increase in the GTP-dependent step in hormonal activation of adenylate cyclase, inasmuch as GTP- and GTP plus hormone-stimulated adenylate cyclase activities were enhanced twofold to threefold in crude membranes prepared from nitrosourea-treated cells. Fluoride-stimulated adenylate cyclase activity was increased only 10--25%. Nicotinamide did not prevent the elevated response, and NAD+ plus NADH levels were not appreciably altered after 42 hours; treatment with streptozotocin. The results suggest a possible involvement of cAMP in the biologic actions of nitrosoureas.