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PMID: 8754823 Published · ppublish English Journal Article Research Support, U.S. Gov't, P.H.S.

Involvement of the transcription factor NF-kappaB in tubular morphogenesis of human microvascular endothelial cells by oxidative stress.

Molecular and cellular biology ·Vol. 16 ·No. 8 ·1996-08-00 ·Pages 4231-9

Shono T, Ono M, Izumi H, Jimi SI, Matsushima K, Okamoto T, Kohno K, Kuwano M

Abstract

Oxygen radicals are induced under various pathologic conditions associated with neovascularization. Oxygen radicals modulate angiogenesis in cultured human microvascular endothelial cells by an unknown mechanism. Treatment of human microvascular endothelial cells for 15 min with 0.1 to 0.5 mM hydrogen peroxide (H2O2) or 100 U of tumor necrosis factor alpha per ml induced tubular morphogenesis in type I collagen gels. Gel shift assays with nuclear extracts demonstrated that H2O2 increases the binding activities of two transcription factors, NF-kappaB and AP-1, but not of Spl. Tumor necrosis factor alpha increased the binding activities of all three factors. A supershift assay with specific antibodies against JunB, JunD, and c-Jun (Jun family) showed that the antibody against c-Jun supershifted the AP-1 complex after H2O2 treatment. Coadministration of the antisense sequence of NF-kappaB inhibited H2O2-dependent tubular morphogenesis, and the antisense c-Jun oligonucleotide caused partial inhibition. The angiogenic factor responsible for H2O2-induced tubular morphogenesis was examined. Cellular mRNA levels of vascular endothelial growth factor and interleukin-8 (IL-8), but not those of transforming growth factor alpha, were increased after treatment with 0.5 mM H2O2. Coadministration of anti-IL-8 antibody inhibited tubular morphogenesis enhanced by H2O2, and IL-8 itself also enhanced the formation of tube-like structures. Treatment with antisense NF-kappaB oligonucleotide completely blocked H2O2-dependent IL-8 production by endothelial cells. The tubular morphogenesis of vascular endothelial cells after treatment with oxidative stimuli and its possible association with NF-kappaB and IL-8, is examined.

MeSH Terms
Base Sequence Cells, Cultured DNA Primers/chemistry DNA, Antisense/pharmacology Endothelium, Vascular/physiology Gene Expression Regulation, Developmental Humans Interleukin-8/physiology Microcirculation/cytology Molecular Sequence Data Morphogenesis NF-kappa B/physiology Neovascularization, Physiologic Oxidation-Reduction Oxidative Stress RNA, Messenger/genetics Sp1 Transcription Factor/metabolism Transcription Factor AP-1/metabolism Tumor Necrosis Factor-alpha/pharmacology
Chemicals
DNA Primers DNA, Antisense Interleukin-8 NF-kappa B RNA, Messenger Sp1 Transcription Factor Transcription Factor AP-1 Tumor Necrosis Factor-alpha
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Shono T
Department of Biochemistry, Kyushu Unviersity School of Medicine, Fukuoka, Japan.
Ono M
Izumi H
Jimi S I
Matsushima K
Okamoto T
Kohno K
Kuwano M
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.16.8.4231
Published
1996-08-00
Pages
4231-9
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC231421
Subset
IM
Grants
NCI NIH HHS · CA-14195 · United States
External Links
PubMed source DOI: 10.1128/MCB.16.8.4231 PMC Full Text View journal details
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