NF-κB and C/EBP transcription factor families synergistically function in mouse serum amyloid A gene expression induced by inflammatory cytokines

Mouse serum amyloid A proteins (SAA) are encoded by multiple genes and the expression of these SAA genes is highly induced during inflammation. We demonstrate that the expression of one of SAA genes (SAA3) is induced by interleukin-1 (IL-1), and that other inflammatory cytokines such as IL-6 and leukemia inhibitory factor, while they themselves are without any effects, enhanced IL-1 induced SAA3 gene expression. The results of mutational analysis on the SAA3 promoter indicate that both the NF-KB and C/EBP transcription factor-binding motifs are essential for cytokine-induced SAA3 gene expression in Hep3B cells. To study further roles of NF-KB and C/EBP transcription factor family members in SAA3 gene activation, expression vectors for NF-KB subunits (p50 and p65) and C/EBP family members (C/EBP-α and NFIL-6, also called C/EBP-β) were co-transfected into Hep3B hepatoma and F9 embryonic carcinoma cells. The results show that, while the expression of p65 alone strongly transactivated a SAA3 gene, p50 did not induce a significant transactivation, and NFIL-6 and C/EBP-α induced only a marginal transactivation when expressed alone. However, the co-expression of p50 or p65 with C/EBP family members did result in the efficient induction of SAA3 gene expression, indicating that the synergy between NF-KB and C/EBP transcription factor families is essential for SAA3 gene expression during inflammation.