Interleukin-1/β represses MRP2 gene expression through inactivation of interferon regulatory factor 3 in HepG2 cells

Keiji Hisaeda, Akihiko Inokuchi, Takanori Nakamura, Yukihide Iwamoto, Kimitoshi Kohno, Michihiko Kuwano, Takeshi Uchiumi

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Abstract

The human multidrug resistance protein 2 (MRP2/ABCC2), expressed on the bile canalicular membrane, mediates the multispecific efflux of several organic anions, including conjugates of glucuronate, sulfate, and glutathione. Expression of MRP2 can be altered in response to environmental stimuli such as cholestasis and jaundice. We previously reported that MRP2 mRNA expression levels are decreased in the nontumorous part of hepatitis C virus-infected human liver tissues, and that inflammatory cytokines inhibit MRP2 expression in human hepatic (HepG2) cells. We investigated the molecular mechanisms by which inflammatory cytokines modulate MRP2 gene expression in hepatic cells. Treatment of human hepatic cells with interleukin-1β (IL-1β) or tumor necrosis factor α resulted in a decrease in the protein and mRNA levels of MRP2. IL-1β inhibited the transcriptional activity of MRP2 promoter constructs by 40%, and this inhibition of MRP2 promoter activity was mediated through the interferon stimulatory response element (ISRE). Electrophoretic mobility shift assays with IL-1β-treated nuclear extracts showed a decrease in the formation of DNA protein complexes, specifically those including interferon regulatory factor 3 (IRF3). Expression of recombinant human IRF3 increased MRP2 promoter activity. Treatment with a specific extracellular signal-regulated kinase inhibitor relieved IL-1β-induced MRP2 mRNA downregulation and abrogated the binding of IRF3 to the ISRE element. In conclusion, IL-1β induces downregulation of the MRP2 gene by inactivating IRF3 binding to ISRE on the MRP2 promoter in human hepatic cells; this inactivation is accomplished via interference with the extracellular signal-regulated kinase pathway.

Original languageEnglish
Pages (from-to)1574-1582
Number of pages9
JournalHepatology
Volume39
Issue number6
DOIs
Publication statusPublished - Jun 1 2004

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Interferon Regulatory Factor-3
Hep G2 Cells
Interleukin-1
Gene Expression
Hepatocytes
Response Elements
Interferons
Extracellular Signal-Regulated MAP Kinases
Messenger RNA
Down-Regulation
Cytokines
Glucuronic Acid
Cholestasis
Electrophoretic Mobility Shift Assay
Jaundice
Bile
Hepacivirus
Sulfates
Anions
Glutathione

All Science Journal Classification (ASJC) codes

  • Hepatology

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Interleukin-1/β represses MRP2 gene expression through inactivation of interferon regulatory factor 3 in HepG2 cells. / Hisaeda, Keiji; Inokuchi, Akihiko; Nakamura, Takanori; Iwamoto, Yukihide; Kohno, Kimitoshi; Kuwano, Michihiko; Uchiumi, Takeshi.

In: Hepatology, Vol. 39, No. 6, 01.06.2004, p. 1574-1582.

Research output: Contribution to journalArticle

Hisaeda, Keiji ; Inokuchi, Akihiko ; Nakamura, Takanori ; Iwamoto, Yukihide ; Kohno, Kimitoshi ; Kuwano, Michihiko ; Uchiumi, Takeshi. / Interleukin-1/β represses MRP2 gene expression through inactivation of interferon regulatory factor 3 in HepG2 cells. In: Hepatology. 2004 ; Vol. 39, No. 6. pp. 1574-1582.
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AU - Kuwano, Michihiko

AU - Uchiumi, Takeshi

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AB - The human multidrug resistance protein 2 (MRP2/ABCC2), expressed on the bile canalicular membrane, mediates the multispecific efflux of several organic anions, including conjugates of glucuronate, sulfate, and glutathione. Expression of MRP2 can be altered in response to environmental stimuli such as cholestasis and jaundice. We previously reported that MRP2 mRNA expression levels are decreased in the nontumorous part of hepatitis C virus-infected human liver tissues, and that inflammatory cytokines inhibit MRP2 expression in human hepatic (HepG2) cells. We investigated the molecular mechanisms by which inflammatory cytokines modulate MRP2 gene expression in hepatic cells. Treatment of human hepatic cells with interleukin-1β (IL-1β) or tumor necrosis factor α resulted in a decrease in the protein and mRNA levels of MRP2. IL-1β inhibited the transcriptional activity of MRP2 promoter constructs by 40%, and this inhibition of MRP2 promoter activity was mediated through the interferon stimulatory response element (ISRE). Electrophoretic mobility shift assays with IL-1β-treated nuclear extracts showed a decrease in the formation of DNA protein complexes, specifically those including interferon regulatory factor 3 (IRF3). Expression of recombinant human IRF3 increased MRP2 promoter activity. Treatment with a specific extracellular signal-regulated kinase inhibitor relieved IL-1β-induced MRP2 mRNA downregulation and abrogated the binding of IRF3 to the ISRE element. In conclusion, IL-1β induces downregulation of the MRP2 gene by inactivating IRF3 binding to ISRE on the MRP2 promoter in human hepatic cells; this inactivation is accomplished via interference with the extracellular signal-regulated kinase pathway.

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