The role of oxygen-derived free radicals and nitric oxide in cytokine-induced antiproliferation of pancreatic cancer cells

Pancreatic cancer cells are susceptible to antiproliferative effects of cytokines such as tumor necrosis factor-alpha (TNF-alpha). However, little is known about the mechanisms involved. To determine the mechanisms of the antiproliferative effects of TNF-alpha on pancreatic cancer cells. In the current study, four of five pancreatic cancer cell lines tested were responsive to the antiproliferative effects of TNF-alpha. In two cell lines, the effects of TNF-alpha were completely abolished by superoxide dismutase, suggesting that superoxide anion mediates the effects. Further, inhibition of nitric oxide (NO) synthase by L-NAME potentiated the TNF-alpha response, suggesting a protective role for endogenously produced NO in these two cell lines. MiaPaCa-2 cells, which were unresponsive to the antiproliferative effects of TNF-alpha, produced five times more nitrite than the other cell lines. Treatment of MiaPaCa-2 cells with interferon-gamma (IFN-gamma) increased inducible nitric oxide synthase mRNA as shown by reverse transcription-polymerase chain reaction. This induction was potentiated by TNF-alpha and further enhanced by a combination of three cytokines (INF-gamma, TNF-alpha, and interleukin 1-beta (IL-1beta). This combination of cytokines increased nitrite accumulation fourfold and inhibited the proliferation of this resistant cell line. These effects were prevented by L-NAME. These studies suggest that TNF-alpha inhibits proliferation of pancreatic cancer cells by increasing the production of superoxide anion and that endogenously produced NO protects against this effect. In addition, there is a direct correlation between the amount of NO produced and resistance to TNF-alpha in the only resistant cell line. In contrast, treatment with a combination of IFN-gamma, TNF-alpha and IL-1beta upregulates inducible nitric oxide synthase, and the resulting markedly enhanced NO production inhibits pancreatic cancer cell growth. This pathway may provide a valuable target for therapy of this devastating disease.