Activation of Nuclear Factor κB and Oncogene Expression by 12(R)-Hydroxyeicosatrienoic Acid, an Angiogenic Factor in Microvessel Endothelial Cells

Michal Laniado-Schwartzman, Yan Lavrovsky, Robert A. Stoltz, Michael S. Conners, John R. Falck, Kamlesh Chauhan, Nader G. Abraham

Research output: Contribution to journalArticle

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Abstract

12(R)-Hydroxy-5,8,14(Z,Z,Z)-eicosatrienoic acid (12(R)-HETrE) is an arachidonic acid metabolite formed by the corneal epithelium of several species, porcine leukocytes, and human and rat epidermal cells. It is a potent, stereospecific proinflammatory and angiogenic factor and its synthesis is increased manyfold in inflamed tissues, e.g. cornea and skin. It is possible that the angiogenic activity of 12(R)-HETrE is due to a direct mitogenic effect on microvessel endothelial cells via yet to be elucidated cellular and molecular mechanisms. In the present study, we demonstrated the ability of 12(R)-HETrE to stimulate the growth of quiescent endothelial cells in a time- and concentration-dependent manner with a maximal effect at 0.1 nM. This effect was highly stereospecific since its enantiomer, 12(S)-HETrE, had no effect within the same concentration range. Northern blot analysis and transient transfection experiments with chloramphenicol acetyltransferase constructs of oncogene promoter regions demonstrated significant increases over control (0.5% fetal calf serum) in c-myc, c-jun, and c-fos mRNA levels and expression in cells treated with 0.1 nM 12(R)-HETrE. Electrophoretic mobility shift assay of nuclear protein extracts from cells treated with 12(R)-HETrE with specific radiolabeled oligonucleotides corresponding to known transcriptional binding sites, including AP-1, AP-2, SP1, TRE, NFκB, TFIID, OKT1, CREB, CTF/NF1, and GRE demonstrated a markedly rapid and specific increase in the binding activity of NFκB and to a lesser extent, AP-1. No significant increase was observed in the binding of other transcription factors assayed as compared to control (untreated) cells. Since the protooncogenes (c-fos, c-jun, and c-myc) are immediate early response genes that are implicated in the process of cell proliferation and differentiation, and activation of certain transcription factors, in particular NFκB, is associated with the immediate response of the cell to an injury, we propose that 12(R)HETrE's mitogenic and angiogenic activities are mediated, in part, via the activation of NFκB and expression of these protooncogenes.

Original languageEnglish (US)
Pages (from-to)24321-24327
Number of pages7
JournalJournal of Biological Chemistry
Volume269
Issue number39
StatePublished - Sep 30 1994

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Angiogenesis Inducing Agents
Endothelial cells
Microvessels
Oncogenes
Endothelial Cells
Chemical activation
Acids
Transcription Factor AP-1
Transcription Factors
Transcription Factor TFIID
Electrophoretic mobility
Corneal Epithelium
Chloramphenicol O-Acetyltransferase
Immediate-Early Genes
Enantiomers
Cell proliferation
Electrophoretic Mobility Shift Assay
Metabolites
Nuclear Proteins
Cell Extracts

ASJC Scopus subject areas

  • Biochemistry

Cite this

Laniado-Schwartzman, M., Lavrovsky, Y., Stoltz, R. A., Conners, M. S., Falck, J. R., Chauhan, K., & Abraham, N. G. (1994). Activation of Nuclear Factor κB and Oncogene Expression by 12(R)-Hydroxyeicosatrienoic Acid, an Angiogenic Factor in Microvessel Endothelial Cells. Journal of Biological Chemistry, 269(39), 24321-24327.

Activation of Nuclear Factor κB and Oncogene Expression by 12(R)-Hydroxyeicosatrienoic Acid, an Angiogenic Factor in Microvessel Endothelial Cells. / Laniado-Schwartzman, Michal; Lavrovsky, Yan; Stoltz, Robert A.; Conners, Michael S.; Falck, John R.; Chauhan, Kamlesh; Abraham, Nader G.

In: Journal of Biological Chemistry, Vol. 269, No. 39, 30.09.1994, p. 24321-24327.

Research output: Contribution to journalArticle

Laniado-Schwartzman, M, Lavrovsky, Y, Stoltz, RA, Conners, MS, Falck, JR, Chauhan, K & Abraham, NG 1994, 'Activation of Nuclear Factor κB and Oncogene Expression by 12(R)-Hydroxyeicosatrienoic Acid, an Angiogenic Factor in Microvessel Endothelial Cells', Journal of Biological Chemistry, vol. 269, no. 39, pp. 24321-24327.
Laniado-Schwartzman, Michal ; Lavrovsky, Yan ; Stoltz, Robert A. ; Conners, Michael S. ; Falck, John R. ; Chauhan, Kamlesh ; Abraham, Nader G. / Activation of Nuclear Factor κB and Oncogene Expression by 12(R)-Hydroxyeicosatrienoic Acid, an Angiogenic Factor in Microvessel Endothelial Cells. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 39. pp. 24321-24327.
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abstract = "12(R)-Hydroxy-5,8,14(Z,Z,Z)-eicosatrienoic acid (12(R)-HETrE) is an arachidonic acid metabolite formed by the corneal epithelium of several species, porcine leukocytes, and human and rat epidermal cells. It is a potent, stereospecific proinflammatory and angiogenic factor and its synthesis is increased manyfold in inflamed tissues, e.g. cornea and skin. It is possible that the angiogenic activity of 12(R)-HETrE is due to a direct mitogenic effect on microvessel endothelial cells via yet to be elucidated cellular and molecular mechanisms. In the present study, we demonstrated the ability of 12(R)-HETrE to stimulate the growth of quiescent endothelial cells in a time- and concentration-dependent manner with a maximal effect at 0.1 nM. This effect was highly stereospecific since its enantiomer, 12(S)-HETrE, had no effect within the same concentration range. Northern blot analysis and transient transfection experiments with chloramphenicol acetyltransferase constructs of oncogene promoter regions demonstrated significant increases over control (0.5{\%} fetal calf serum) in c-myc, c-jun, and c-fos mRNA levels and expression in cells treated with 0.1 nM 12(R)-HETrE. Electrophoretic mobility shift assay of nuclear protein extracts from cells treated with 12(R)-HETrE with specific radiolabeled oligonucleotides corresponding to known transcriptional binding sites, including AP-1, AP-2, SP1, TRE, NFκB, TFIID, OKT1, CREB, CTF/NF1, and GRE demonstrated a markedly rapid and specific increase in the binding activity of NFκB and to a lesser extent, AP-1. No significant increase was observed in the binding of other transcription factors assayed as compared to control (untreated) cells. Since the protooncogenes (c-fos, c-jun, and c-myc) are immediate early response genes that are implicated in the process of cell proliferation and differentiation, and activation of certain transcription factors, in particular NFκB, is associated with the immediate response of the cell to an injury, we propose that 12(R)HETrE's mitogenic and angiogenic activities are mediated, in part, via the activation of NFκB and expression of these protooncogenes.",
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N2 - 12(R)-Hydroxy-5,8,14(Z,Z,Z)-eicosatrienoic acid (12(R)-HETrE) is an arachidonic acid metabolite formed by the corneal epithelium of several species, porcine leukocytes, and human and rat epidermal cells. It is a potent, stereospecific proinflammatory and angiogenic factor and its synthesis is increased manyfold in inflamed tissues, e.g. cornea and skin. It is possible that the angiogenic activity of 12(R)-HETrE is due to a direct mitogenic effect on microvessel endothelial cells via yet to be elucidated cellular and molecular mechanisms. In the present study, we demonstrated the ability of 12(R)-HETrE to stimulate the growth of quiescent endothelial cells in a time- and concentration-dependent manner with a maximal effect at 0.1 nM. This effect was highly stereospecific since its enantiomer, 12(S)-HETrE, had no effect within the same concentration range. Northern blot analysis and transient transfection experiments with chloramphenicol acetyltransferase constructs of oncogene promoter regions demonstrated significant increases over control (0.5% fetal calf serum) in c-myc, c-jun, and c-fos mRNA levels and expression in cells treated with 0.1 nM 12(R)-HETrE. Electrophoretic mobility shift assay of nuclear protein extracts from cells treated with 12(R)-HETrE with specific radiolabeled oligonucleotides corresponding to known transcriptional binding sites, including AP-1, AP-2, SP1, TRE, NFκB, TFIID, OKT1, CREB, CTF/NF1, and GRE demonstrated a markedly rapid and specific increase in the binding activity of NFκB and to a lesser extent, AP-1. No significant increase was observed in the binding of other transcription factors assayed as compared to control (untreated) cells. Since the protooncogenes (c-fos, c-jun, and c-myc) are immediate early response genes that are implicated in the process of cell proliferation and differentiation, and activation of certain transcription factors, in particular NFκB, is associated with the immediate response of the cell to an injury, we propose that 12(R)HETrE's mitogenic and angiogenic activities are mediated, in part, via the activation of NFκB and expression of these protooncogenes.

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