RESUMEN
We have previously shown that administration of a low-dose of melphalan (L-phenylalanine mustard; L-PAM) to mice bearing a large s.c. MOPC-315 tumor leads to up-regulation of TNF-alpha expression, which is first evident at the mRNA level at 24 h after the chemotherapy. In this study, we show accumulation of IFN-beta mRNA in the spleen and tumor nodule of such mice as early as 1 h after the chemotherapy followed by elevated production of IFN-beta protein. IFN-beta protein in turn was found to be important for the L-PAM-induced up-regulation of TNF-alpha expression, as neutralization of IFN-beta inhibited the L-PAM-induced up-regulation of TNF-alpha mRNA expression in MOPC-315 tumor cells. In addition, L-PAM failed to up-regulate TNF-alpha expression in spleen cells from mice in which signaling by IFN-beta is deficient. Studies into the mechanism through which L-PAM leads to rapid accumulation of IFN-beta mRNA revealed that it requires de novo RNA synthesis, indicating that the regulation is at the transcriptional level. However, it did not require de novo protein synthesis, indicating that activation of pre-existing transcription factors is sufficient for IFN-beta gene expression. The L-PAM-induced accumulation of IFN-beta mRNA was mimicked with H(2)O(2) and was prevented with the antioxidant N-acetyl-L-cysteine, indicating that reactive oxygen species are involved in the transcriptional regulation of L-PAM-induced IFN-beta gene expression. Thus, the IFN-beta gene is an early response gene that is activated in response to L-PAM via a pathway that involves reactive oxygen species, and IFN-beta in turn plays an important role in L-PAM-induced TNF-alpha up-regulation.
