Stimulation of the DNA binding activity of AP-1 by the peroxisome proliferator ciprofibrate and eicosanoids in cultured rat hepatocytes

Abstract

Peroxisome proliferators induce hepatic peroxisome proliferation and hepatic tumors in rodents. These chemicals increase the expression of the peroxisomal β-oxidation pathway and the cytochrome P-450 4A family, which metabolize lipids, including eicosanoids. Peroxisome proliferators also induce increased cell proliferation in vivo. However, peroxisome proliferators are only weakly mitogenic and are not comitogenic with epidermal growth factor (EGF) in cultured hepatocytes. Our earlier studies found that the peroxisome proliferator ciprofibrate is comitogenic with eicosanoids. We therefore hypothesized that the comitogenicity of the peroxisome proliferator ciprofibrate and eicosanoids may result from a synergistic increase of the DNA binding activity of AP-1. Primary rat hepatocytes were cultured on collagen gels in serum-free L-15 medium with ciprofibrate, eicosanoids, and/or growth factors. The DNA binding activity of AP-1 was determined in nuclear protein extracts by electrophoretic mobility shift assay. The DNA binding activity of AP-1 was not induced by ciprofibrate or eicosanoids alone, but the addition of eicosanoids along with ciprofibrate increased the induction of DNA binding activity of AP-1 at 30 min and 2 h after exposure. The combination of ciprofibrate and PGF 2 α blocked the inhibitory effect of transforming growth factor (TGF)- β on the DNA binding activity of AP-1 induced by EGF. These results show that the peroxisome proliferator ciprofibrate and eicosanoids co-stimulate the DNA binding activity of AP-1 and suggest that changes in eicosanoid concentrations may modulate mitogenic signal transduction pathways by the peroxisome proliferator ciprofibrate.