Transcription factor decoy for nuclear factor-κB inhibits tumor necrosis factor-α-induced expression of interleukin-6 and intracellular adhesion molecule-1 in endothelial cells

Tomita, Naruya; Morishita, Ryuichi; Tomita, Sayuri; Yamamoto, Kei; Aoki, Motokuni; Matsushita, Hisashi; Hayashi, Shinichiro; Higaki, Jitsuo; Ogihara, Toshio

doi:10.1097/00004872-199816070-00013

Cited by 64 publications

(44 citation statements)

References 30 publications

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“…HCAEC were transfected with double-stranded phosphorothioate oligonucleotides (2 mol/L) containing either the HSE consensus sequence 5ЈGCCTCGAATG-TTCGCGAACTTT3Ј or NF-B 5ЈCCTTGAAGGGATTTC-CCTCC3Ј (Trilink, San Diego, Calif) using 5 g/ L Lipofectin (Gibco BRL, Carlsbad, Calif), which is routinely used to facilitate efficient uptake of the oligonucleotide decoys by vascular cells. 17 Twelve hours later, 17␤-estradiol was added to the media to a final concentration of 100 nM. A scrambled double-stranded phosphorothioate oligonucleotide with the sequence 5ЈATGGCCTCGTTTAT-TCACGCG3Ј was used as a control.…”

Section: Transcription Factor Decoy Experimentsmentioning

confidence: 99%

Estrogen, Heat Shock Proteins, and NFκB in Human Vascular Endothelium

Hamilton

Mbai

Gupta

et al. 2004

ATVB

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Background-We hypothesized that estrogen would increase HSP72 in human coronary artery endothelial cells (HCAEC), and that these would be more sensitive to estrogen than our previous observations in myocytes. Methods and Results-HCAEC were treated with 17␤-estradiol or tamoxifen, ranging from physiological to pharmacological(1 nM to 10 mol/L) for either 24 hours (early) or 7 days (chronic). HSP expression was assessed by Western blots. Both early and chronic 17␤-estradiol and tamoxifen increased HSP72. Electromobility shift assays (EMSA) showed activation of HSF-1 with early, but not chronic, 17␤-estradiol. 17␤-Estradiol activated NF B within 10 minutes, and the ER-␣ selective inhibitor, ICI 182 780, abolished this effect. Transcription factor decoys containing the heat shock element blocked HSP72 induction. Estrogen pretreatment decreased lactate dehydrogenase release with hypoxia. This protective effect persisted despite blockade of HSF-1 by decoys. However, an NF-B decoy prevented the increase in HSP72 and abolished the estrogen-associated protection during hypoxia. Conclusions-17␤-Estradiol upregulates HSP72 early and chronically via different mechanisms in HCAEC, and provides cytoprotection during hypoxia, independent of HSP72 induction. NF-B mediates the early increase in HSP72, suggesting that estrogen activates NF-B via a nongenomic, receptor-dependent mechanism, and this leads to activation of HSF-1. Activation of NF-B was critical for estrogen-associated protection. Further studies are needed to elucidate the involved signaling pathways.

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Section: Transcription Factor Decoy Experimentsmentioning

confidence: 99%

Estrogen, Heat Shock Proteins, and NFκB in Human Vascular Endothelium

Hamilton

Mbai

Gupta

et al. 2004

ATVB

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“…The ODN were resuspended in sterile water to a concentration of 1.6 mg/ml and the forward and reverse ODN were annealed over 2 h using a temperature gradient ranging from 80°C to 25°C as previously published. 40 Gene Therapy …”

Section: Nf B Decoy Oligonucleotidesmentioning

confidence: 99%

Cytoplasmic deposition of NFκB decoy oligonucleotides is insufficient to inhibit bleomycin-induced pulmonary inflammation

et al. 2002

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Lung inflammation leads to severe tissue destruction and ultimately organ failure in a number of diseases, including cystic fibrosis (CF). The transcription factor nuclear factor kappa B (NF B) regulates expression of many pro-inflammatory mediators. We have assessed the effect of topical administration of NF B decoys in a bleomycin model of acute lung inflammation. Using fluorescein-labelled decoy oligonucleotides (ODN) (80 g/mouse) we have shown that lipid-complexed and 'naked' ODN transfect conducting airway epithelium in a comparable manner (approximately 65% of cells). However, the ODN were detectable in the cytoplasm, but not in the nucleus of transfected cells. An increase of ODN dose to 500 g/mouse did not increase

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“…[15][16][17][18] In addition, synthetic double-stranded DNA (dsDNA) with high affinity for NFkB, NFkB decoy, have been shown to block the induction of gene expression mediated by NFkB in vitro 15,[19][20][21][22] and in vivo. 23,24 With the use of decoy oligonucleotides, the function of a particular DNA-binding protein or protein complex can be inhibited since the decoy competes for protein binding with the authentic binding elements, interfering with gene regulation.…”

Section: Introductionmentioning

confidence: 99%

Cellular delivery of a double-stranded oligonucleotide

et al. 2004

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The activation of nuclear factor kB (NFkB) is a key event in immune and inflammatory responses. In this study, a cellpenetrating transport peptide, transportan (TP) or its shorter analogue TP 10, was used to facilitate the cellular uptake of an NFkB decoy. Peptide nucleic acid (PNA) hexamer or nonamer was linked to the transport peptide by a disulfide bond. NFkB decoy oligonucleotide consisted of a doublestranded consensus sequence corresponding to the kB site localized in the IL-6 gene promoter, 5 0 -GGGACTTTCCC-3 0 , with a single-stranded protruding 3 0 -terminal sequence complementary to the PNA sequence was hybridized to the transport peptide-PNA construct. The ability of the transport peptide-PNA-NFkB decoy complex to block the effect of interleukin (IL)-1b-induced NFkB activation and IL-6 gene expression was analyzed by electrophoretic mobility shift assay and reverse transcriptase-polymerase chain reaction in rat Rinm5F insulinoma cells. Preincubation with transport peptide-PNA-NFkB decoy (1 mM, 1 h) blocked IL-1b-induced NFkB-binding activity and significantly reduced the IL-6 mRNA expression. The same concentration of NFkB decoy in the absence of transport peptide-PNA had no effect even after longer incubations. Our results showed that binding of the oligonucleotide NFkB decoy to the nonamer PNA sequence resulted in a stable complex that was efficiently translocated across the plasma membrane.

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Transcription factor decoy for nuclear factor-κB inhibits tumor necrosis factor-α-induced expression of interleukin-6 and intracellular adhesion molecule-1 in endothelial cells

Abstract: Our results demonstrate that nuclear factor-kappaB decoy oligodeoxynucleotides transfected by cationic liposome method inhibited tumor necrosis factor--induced expression of interleukin-6 and intracellular adhesion molecule-1 in endothelial cells.

Cited by 64 publications

References 30 publications

Estrogen, Heat Shock Proteins, and NFκB in Human Vascular Endothelium

Estrogen, Heat Shock Proteins, and NFκB in Human Vascular Endothelium

Cytoplasmic deposition of NFκB decoy oligonucleotides is insufficient to inhibit bleomycin-induced pulmonary inflammation

Cellular delivery of a double-stranded oligonucleotide

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