Regulation of intracellular polyamine biosynthesis and transport by NO and cytokines TNF-alpha and IFN-gamma

Nitric oxide (NO)has been described to exert cytostatic effects on cellularproliferation; however the mechanisms responsible for these effectshave yet to be fully resolved. Polyamines, conversely, are requiredcomponents of cellular proliferation. In experimental models ofinflammation, a relationship between these two pathways has beensuggested by the temporal regulation of a common precursor, arginine.This study was undertaken to determine the effects NO and the NOsynthase (NOS)-inducing cytokines, tumor necrosis factor- (TNF-)and interferon- (IFN-), exert on polyamine regulation. Thetransformed kidney proximal tubule cell line, MCT, maintains highconstitutive levels of the first polyamine biosynthetic enzyme, ornithine decarboxylase (ODC). NO donors markedly suppressed ODC activity in MCT and all other cell lines examined. TNF- and IFN- induction of NO generation resulted in suppressed ODC activity, aneffect prevented by the inducible NOS inhibitorL-N⁶-(1-iminoethyl)lysine(L-NIL). Dithiothreitol reversalof NO-mediated ODC suppression supports nitrosylation as the mechanismof inactivation. We also evaluated polyamine uptake, inasmuch asinhibition of ODC can result in a compensatory induction of polyaminetransporters. Administration of NO donors, or TNF- and IFN-,suppressed³H]putrescine uptake,thereby preventing transport-mediated reestablishment of intracellularpolyamine levels. This study demonstrates the capacity of NO andinflammatory cytokines to regulate both polyamine biosynthesis and transport.