CONTROL OF ACTIVATOR PROTEIN-1 AND NUCLEAR FACTOR-KAPPA-B ACTIVITY BY INTERLEUKIN-1, INTERLEUKIN-6 AND METALS IN HEPG2 CELLS

The intracellular signals induced by IL-1 and IL-6 have been described but there are few details of the signals they induce in liver-derived cells during initiation of acute phase protein synthesis. We therefore used an in vitro system to investigate signalling by IL-1 and IL-6 in the human liver cell line, HepG2. Chloramphenicol acetyl transferase (CAT) expression vectors, under the control of activator protein-1 (pTRE-CAT), nuclear factor kappa B (pNF-CAT) or no enhancer region (pBLCAT2), were transfected into HepG2 cells and the effects of the cytokines on their activity was studied. Profound changes in liver processing of heavy metals and the induction of metal-dependent acute proteins are also seen during the acute phase response. To determine if the supply of metal ions could itself influence signalling we also investigated the effects of cadmium and zinc on the activity of the transfected vectors. Both alpha and beta forms of interleukin-1 increased the expression of pTRE-CAT and pNF-CAT, but not pBLCAT2, while interleukin-6 had no effect, suggesting that activator protein-1 and nuclear factor kappa B activity was induced by interleukin-1, but not interleukin-6. Specificity of the effect of interleukin-1 alpha was confirmed using an anti-interleukin-1 alpha monoclonal antibody. Zinc and cadmium also increased pTRE-CAT expression, but not pNF-CAT or pBLCAT2. Removal of heavy metal ions from the culture medium resulted in decreased pTRE-CAT expression, while pNF-CAT and pBLCAT2 were relatively unaffected, confirming the stimulatory effect of metals on activator protein-1, but not nuclear protein kappa B activity. Therefore, metal and interleukin-1-mediated signal transduction may involve overlapping pathways, whereas interleukin-l and interleukin-6 act via different pathways in liver cells.