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

168 Citations (Scopus)

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)
JournalAmerican Journal of Physiology - Gastrointestinal and Liver Physiology
Volume266
Issue number5 29-5
StatePublished - 1994
Externally publishedYes

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Prostaglandin-Endoperoxide Synthases
Organism Cloning
Intercellular Signaling Peptides and Proteins
Epithelial Cells
COS Cells
Transforming Growth Factors
Mitogens
Genes
Messenger RNA
Eicosanoids
Cyclooxygenase 2
Prostaglandins
Proteins
Complementary DNA
Kidney
Amino Acids
Brain
Growth

Keywords

  • eicosanoids
  • proliferation
  • transforming growth factor-α

ASJC Scopus subject areas

  • Physiology
  • Gastroenterology

Cite this

Cloning and characterization of a growth factor-inducible cyclooxygenase gene from rat intestinal epithelial cells. / DuBois, R. N.; Tsujii, M.; Bishop, P.; Awad, J. A.; Makita, K.; Lanahan, A.

In: American Journal of Physiology - Gastrointestinal and Liver Physiology, Vol. 266, No. 5 29-5, 1994.

Research output: Contribution to journalArticle

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AU - Tsujii, M.

AU - Bishop, P.

AU - Awad, J. A.

AU - Makita, K.

AU - Lanahan, A.

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N2 - 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.

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