Nitric oxide is produced during TNF killing of U937A cells but does not contribute to the cytotoxic process.

Proinflammatory cytokines, including TNF, stimulate nitric-oxide free radical production in a variety of tissues through the induction of the enzyme nitric-oxide synthase. As free radicals are considered likely candidates in the cytotoxic action of TNF, we examined nitric oxide production in TNF-sensitive U937A and TNF-resistant U937A/R cells and its potential role in TNF-induced cytotoxicity. TNF stimulated U937A nitrite production through a process that was abolished by the competitive inhibitors of nitric-oxide synthase N-omega-nitro-L-arginine methyl ester (NAME) and N-G-monomethyl-L-arginine (NMMA) without inhibition of TNF-induced cytotoxicity. TNF also increased nitrite production in TNF-resistant U937A/R cells. In addition, the cytotoxic action of TNF was independent of L-arginine substrate availability. Thus, although cytokine-inducible nitric oxide production is emerging as an effective antitumour mechanism, here, TNF clearly exerted potent antitumour activity against U937A cells through a nitric-oxide-independent mechanism.