Abstract
PURPOSE: The precise molecular targets of interferon-alpha (IFN-alpha) therapy of melanoma are unknown but likely involve signal transducer and activator of transcription 1 (STAT1) signal transduction within host immune effector cells. We hypothesized that microarray analysis could be utilized to identify candidate molecular targets important for mediating the anti-tumor effect of exogenously administered IFN-alpha. EXPERIMENTAL METHODS: To identify the STAT1-dependent genes regulated by IFN-alpha, the gene expression profile of splenocytes from wild type (WT) and STAT1(-/-) mice was characterized. RESULTS: This analysis identified 30 genes that required STAT1 signal transduction for optimal expression in response to IFN-alpha (p < 0.001). These genes include granzyme b (Gzmb), interferon regulatory factor 7 (Irf7), Fas death domain-associated protein (Daxx), and lymphocyte antigen 6 complex, locus C (Ly6c). The expression of 20 genes was found to be suppressed in the presence of STAT1 including chemokine ligand 2 (Ccl2), Ccl5, and Ccl7. Nineteen genes were significantly upregulated in murine splenocytes following treatment with IFN-alpha regardless of the presence of STAT1 including CD86, lymphocyte antigen 6 complex, locus A (Ly6a), and Tap binding protein (Tapbp). The expression of representative IFN-responsive genes was confirmed at the transcriptional level by Real Time PCR. CONCLUSION: This report is the first to demonstrate that STAT1-mediated signal transduction plays a major role in the transcriptional response of murine immune cells to IFNalpha.