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Papers In Press, published online ahead of print April 26, 2002
Department of Pharmacology and Toxicology, Dartmouth Medical School, Hanover, NH 03755-3835
Corresponding Author: barchowsky{at}dartmouth.edu
Inhalation of particulate nickel subsulfide (Ni3S2) causes chronic active inflammation and fibrosis of the lungs. However, the mechanisms for these effects are not well understood. Therefore, cell culture experiments with BEAS-2B human airway epithelial cells were conducted to test the hypothesis that exposure to non-cytotoxic levels of Ni3S2 induces expression of inflammatory cytokines, such as interleukin-8 (IL-8). Exposure to Ni3S2 for 48h was required to significantly increase IL-8 protein levels. Transcriptional stimulation of IL-8 mRNA levels preceded the increase in protein. Transient exposure to soluble nickel sulfate failed to increase IL-8 mRNA. Transfection with truncated IL-8 promoter constructs linked to the luciferase gene demonstrated that nickel-induced IL-8 transcription required –272 bp of the promoter relative to the transcriptional start site. A –133 bp construct, containing cytokine and hypoxia-sensitive AP-1, NF-IL6 and NF-
J. Biol. Chem, 10.1074/jbc.M202941200
Submitted on March 27, 2002
Revised on April 26, 2002
Accepted on April 26, 2002
Novel pathway for nickel-induced interleukin-8 expression
B sites, was insufficient for induction by nickel. Transfection with a dominant negative AP-1 construct or mutation of the AP-1, GATA, or C/EBP sites in the -272 bp IL-8 promoter construct blocked induction by nickel. Inhibiting ERK, phosphatidylinositol 3-kinase, but not p38 kinase, diacylglycerol kinase or hypoxia inducible factor-1
, attenuated nickel induction of IL-8. These studies indicate that nickel induced IL-8 transcription through a novel pathway that requires both AP-1 and non-traditional transcription factors.
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