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Vol. 30, Issue 11, 1186-1193, November 2002
in Human
Intestinal Epithelial Cells (Caco-2) in Culture
Department of Biomedical Sciences, University of Guelph, Guelph,
Ontario, Canada
The influence of pro-inflammatory cytokines on alpha class
glutathione S-transferase A1 and A2 (GSTA1/A2)
expression was examined in human colonic epithelial cells (Caco-2) in
culture. Dose-dependent reductions in GSTA1/A2 mRNA, protein, and
activity levels occurred in Caco-2 cells cultured in conditioned medium
(CM) from lipopolysaccharide-stimulated murine monocyte-macrophage
cells (RAW 264.7). Neutralizing anti-interleukin-1
(IL-1)
antibodies attenuated this repression of GSTA1/A2 expression by CM.
Moreover, recombinant human IL-1 reduced GST
expression at the
mRNA, protein, and activity levels in a dose-related fashion. Reduction
of GSTA1/A2 mRNA levels by IL-1 was attenuated by pretreatment with
IL-1 receptor antagonist. GSTA1/A2 mRNA half-lives were similar in
control and IL-1-treated cells, indicating that IL-1 has no
effect on mRNA stability. In reporter gene studies, IL-1 caused a
dose-related reduction of luciferase activity in Caco-2 cells transfected with the full-length GSTA1 promoter-luciferase construct. Using truncated constructs, IL-1 responsiveness was mapped to a
region 286 base pairs upstream to the coding region. Deletion of a
hepatic nuclear factor 1 (HNF-1) site in this region abrogated the
IL-1-mediated repression of GSTA1 promoter activity. These results
demonstrate that IL-1 down-regulates GSTA1/A2 expression in cultured
human enterocytes by a transcriptional mechanism involving an HNF-1 site.
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