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Am J Physiol Lung Cell Mol Physiol 274: L922-L932, 1998;
1040-0605/98 $5.00
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Vol. 274, Issue 6, L922-L932, June 1998

Cytokine-induced nitric oxide formation in normal but not in neoplastic murine lung epithelial cell lines

David C. Thompson1, Stephanie E. Porter1, Alison K. Bauer2, Kumuda C. Das3, Brandon Ou1, Lori Dwyer-Nield1, Carl W. White3, and Alvin M. Malkinson1

Departments of 1 Pharmaceutical Sciences and 2 Pharmacology, University of Colorado Health Sciences Center, Denver 80262-0238; and 3 Department of Pediatrics, National Jewish Center for Immunology and Respiratory Medicine, Denver, Colorado 80206

Cytomix, a mixture of interferon-gamma , tumor necrosis factor-alpha , and interleukin-1beta , induces nitric oxide (NO) production in lung epithelial cell lines. It is not known whether neoplastic transformation alters a cell's ability to form NO in response to cytokines. The present study investigated NO formation in two murine lines of immortalized "normal" (nontumorigenic) lung epithelial cells of alveolar type II origin, E10 and C10, and their sibling spontaneous transformants, E9 and A5. Nontumorigenic cells elaborated much more NO after cytomix exposure than did their tumorigenic counterparts. NO production was prevented by inhibiting protein synthesis and NO synthase and attenuated by dexamethasone. Northern and Western blot analyses of inducible NO synthase (iNOS) demonstrated cytomix-induced induction of iNOS only in nontumorigenic cells. The deficiency in NO production in tumorigenic cells was not associated with reduced iNOS mRNA stability or with differences in cytomix-induced nuclear factor-kappa B activation. Although cytomix caused a greater production of NO in E10 cells than in E9 cells, the same treatment induced equivalent proliferation in both cell lines. These results indicate a specific deficiency in cytokine-induced NO synthesis in transformed murine lung epithelial cells relative to their normal progenitor cells and provide a model for investigating iNOS regulation.

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