Relationship of trichothecene structure to COX-2 induction in the macrophage: selective action of type B (8-keto) trichothecenes.
The trichothecene mycotoxin deoxynivalenol (DON, vomitoxin), when at partially cytotoxic concentrations, induces cyclooxygenase-2 (COX-2) expression by promoting transcriptional activity and mRNA stability via mitogen-activated protein kinase (MAPK) signaling pathways. The purpose of this study was to test the hypothesis that trichothecenes differentially affect COX-2 gene expression and that these effects were related to MAPK activation. Representative members of the three major trichothecene families (A, B, and D) were compared for their capacity to induce COX-2 in the RAW 264.7 murine macrophage cell line. When cells were treated with concentrations that inhibited the 3-(4,5-di-methylthizol-2-yl)-2,5 diphenyl tetrazolium bromide (MTT) viability response by 20% (IC20), Type B trichothecenes including DON, 15-acetyl-DON, 3-acetyl-DON, and fusarenon-X were found to be effective inducers of COX-2 mRNA expression, whereas equitoxic Type A and Type D trichothecenes had markedly less effects. To compare effects of COX-2 gene transactivation and mRNA stabilization, luciferase reporter vectors containing 5'-promoter or 3'-untranslated regions of the gene, respectively, were transfected into RAW 264.7 cells and the effects of various trichothecenes on luciferase activities were measured. Type B but not Type A or D toxins at concentrations up to the MTT IC50 enhanced luciferase activities, indicating preferential COX-2 transcriptional activation and mRNA stabilization by this trichothecene subset. At their respective IC20s, Type B trichothecenes also significantly activated the three major MAPK families, whereas Type A and D did not. Blocking ERK and p38 with chemical inhibitors significantly suppressed Type B-induced COX-2 expression. Although JNK reportedly contributes to COX-2 expression in the other signaling models, transfection with the dominant negative JNK vector did not diminish the COX-2 expression. Taken together, Type B trichothecenes selectively enhanced transcription and stabilization of the COX-2 gene, and this was mediated by the ERK 1/2 and p38 signaling pathways. Selective action on COX-2 might contribute to unique pathologic manifestations associated with Type B trichothecene-mediated immunotoxicity.
D000818:Animals; D002478:Cells, Cultured; D051546:Cyclooxygenase 2; D004790:Enzyme Induction / Q000187:drug effects*; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D020928:Mitogen-Activated Protein Kinases / Q000187:drug effects*; D011451:Prostaglandin-Endoperoxide Synthases / Q000096:biosynthesis* / Q000235:genetics; D013329:Structure-Activity Relationship; D014255:Trichothecenes / Q000633:toxicity*
D007527:Isoenzymes; D014255:Trichothecenes; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases; D020928:Mitogen-Activated Protein Kinases