Characterization of the Pattern of the Nongenomic Signaling Pathway Through Which TCDD-induces Early Inflammatory Responses in U937 Human Macrophages

2,3,7,8-Tetrachlorodibenzo(p)dioxin (TCDD) has been known to induce inflammatory signaling in a number of cell types and tissues. We found that in U937 macrophages TCDD causes rapid activation of cytosolic phospholipase A2 (cPLA2) within 30min as judged by the increase in the serine 505 phosphorylated form of cPLA2 protein and the increased cellular release of free arachidonic acid. This initial action of TCDD is accompanied with the up-regulation of an important inflammation marker, COX-2 mRNA expression within 1h, and by 3h, several other markers become up-regulated. These effects appear to be dependent on the initial increase in the intracellular concentration of Ca(2+), and activation of cPLA2 and COX-2. A comparative study among three different human cell lines showed that activation of COX-2 within 1h of action of TCDD is a common feature exhibited by all cell lines. On the other hand, the U937 macrophage line appears to be unique among them with respect to its ability to activate TNF-alpha and IL-8 mRNA expressions, and not requiring Src kinase in propagating the initial signaling of cPLA2. Based on the rapidity of activation of cPLA2 and COX-2, which occurs within 1h of cell exposure to TCDD, when no change in mRNA expression of CYP1A1 has been observed, it is apparent that this unique action of TCDD is carried out through a distinct "nongenomic" pathway which, is clearly discernable from the classical, "genomic" action pathway of the AhR by not requiring the participation of ARNT.