Regulation of 12-lipoxygenase in rat intestinal epithelial cells during differentiation and apoptosis induced by sodium butyrate

Hideki Kamitani, Hiroshi Ikawa, Linda C. Hsi, Takashi Watanabe, Raymond N. DuBois, Thomas E. Eling

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    29 Scopus citations

    Abstract

    We evaluated the expression and activity of rat 12-lipoxygenase (LO) in rat intestinal epithelial (RIE) cells during apoptosis and cell differentiation. Sodium butyrate (NaBT) treatment induced wild-type RIE (W- RIE) cells to undergo differentiation and apoptosis. Alkaline phosphatase (ALP) activity, a marker of cell differentiation, and DNA fragmentation, an index of apoptosis, were increased by NaBT treatment. Arachidonic acid was metabolized primarily to 12-hydroxyeicosatetraenoic acid (HETE) suggesting induction of 12-LO activity. In contrast, sense-RIE (S-RIE) cells engineered to overexpress COX-2 were resistant to apoptosis by treatment with 5 mM NaBT and NaBT did not induce 12-LO activity. The upregulation of 12-LO expression by NaBT in W-RIE cells was confirmed at both the transcriptional and translational level but 12-LO was undetectable in S-RIE cells following NaBT treatment. The expression of 12-LO mRNA in W-RIE cells occurs as early as 6 h after treatment and reaches maximum expression at 24 h following treatment. This inducible 12-LO was isolated by RT-PCR and identified as rat 'leukocyte- type' 12-LO. The level of 12-LO expression in W-RIE cells was dependent on the concentration of NaBT and appears to reflect the extent of cell differentiation. NDGA, a lipoxygenase inhibitor, attenuated induction of ALP activity by NaBT treatment of W-RIE cells. These observations suggested that 12-LO is regulated by treatment with NaBT and is associated with cell differentiation in rat intestinal epithelial cells.

    Original languageEnglish (US)
    Pages (from-to)45-55
    Number of pages11
    JournalArchives of Biochemistry and Biophysics
    Volume368
    Issue number1
    DOIs
    StatePublished - Aug 1 1999

    Keywords

    • 12-lipoxygenase
    • Alkaline phosphatase
    • Cell differentiation
    • NDGA
    • Sodium butyrate

    ASJC Scopus subject areas

    • Biophysics
    • Biochemistry
    • Molecular Biology

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