Cloning and characterization of a growth factor-inducible cyclooxygenase gene from rat intestinal epithelial cells

R. N. DuBois, M. Tsujii, P. Bishop, J. A. Awad, K. Makita, A. Lanahan

    Research output: Contribution to journalArticle

    170 Scopus citations

    Abstract

    Growth factors have been shown to play a role in intestinal epithelial growth regulation and transformation. Utilizing standard differential cloning techniques, we have isolated a growth factor-inducible gene (RS-2) from rat intestinal epithelial cells that has ~95% homology to the mouse mitogen- inducible cyclooxygenase (COX-2) at the amino acid level. This cDNA hybridizes to a ~4.5-kb mRNA from transforming growth factor (TGF)-α- stimulated rat intestinal epithelial (RIE-1) cells and is constitutively expressed in vivo in adult rat kidney and brain. Nuclear run-on experiments demonstrate that the increase of RS-2 mRNA after TGF-α stimulation is in part due to an increased transcription rate of the gene. The coding region for RS-2 was subcloned into a pCMV-2 expression vector, and the RS-2 protein was expressed in COS-1 cells. Microsomal fractions isolated from the COS-1 cells transfected with the RS-2 expression vector contained cyclooxygenase activity. In addition to the production of prostaglandins, the recombinant RS-2 protein also catalyzed the formation of three other eicosanoid products. In summary, we have cloned a mitogen-inducible cyclooxygenase gene from rat intestinal cells that is induced following growth factor stimulation.

    Original languageEnglish (US)
    Pages (from-to)G822-G827
    JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
    Volume266
    Issue number5 29-5
    DOIs
    StatePublished - 1994

    Keywords

    • eicosanoids
    • proliferation
    • transforming growth factor-α

    ASJC Scopus subject areas

    • Physiology
    • Hepatology
    • Gastroenterology
    • Physiology (medical)

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