Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo

Cristiana Griffoni, Enzo Spisni, Spartaco Santi, Massimo Riccio, Tiziana Guarnieri, Vittorio Tomasi

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Knock-out of the gene coding for caveolin-1, the main organizer of caveolae, has not yet been performed. We devised a strategy to knock-down caveolin-1 gene expression using antisense oligodeoxynucleotides (ODNs). Seven ODNs, covering different regions of caveolin-1 mRNA, were screened by Western blot analysis of caveolin-1 levels. The most active and specific was found to reduce caveolin-1 protein levels by 70% at 1 μM concentration and its action, as demonstrated by a marked reduction (about 50%) in caveolin-1 mRNA levels, was due to a true antisense mechanism. In HUVEC treated with the active ODN, caveolae were undetectable by confocol and electron microscopy, while their number was not affected when cells were treated with a scrambled ODN. Using the fibrin gel 3 D angiogenesis test we established that the active (but not the scrambled) ODN strongly suppressed capillary-like tube formation. Moreover, an antisense tailored against chicken caveolin-1 mRNA, when tested using the chorio-allantoic membrane technique, dramatically reduced vessel formation at doses (10-20 μg) under which control ODNs were ineffective and devoid of toxicity. Thus, it is likely that caveolin-1 down regulation, followed by caveolae disruption, impairs angiogenesis in vitro and in vivo. (C) 2000 Academic Press.

Original languageEnglish
Pages (from-to)756-761
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume276
Issue number2
DOIs
Publication statusPublished - Sep 24 2000

Fingerprint

Caveolin 1
Antisense Oligonucleotides
Oligodeoxyribonucleotides
Caveolae
Messenger RNA
Allantois
Caveolins
Gene Knockout Techniques
In Vitro Techniques
Fibrin
Gene expression
Electron microscopy
Toxicity
Chickens
Electron Microscopy
Down-Regulation
Genes
Western Blotting
Gels
Membranes

Keywords

  • Angiogenesis
  • Antisense oligonucleotides
  • Caveolin

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo. / Griffoni, Cristiana; Spisni, Enzo; Santi, Spartaco; Riccio, Massimo; Guarnieri, Tiziana; Tomasi, Vittorio.

In: Biochemical and Biophysical Research Communications, Vol. 276, No. 2, 24.09.2000, p. 756-761.

Research output: Contribution to journalArticle

Griffoni, Cristiana ; Spisni, Enzo ; Santi, Spartaco ; Riccio, Massimo ; Guarnieri, Tiziana ; Tomasi, Vittorio. / Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo. In: Biochemical and Biophysical Research Communications. 2000 ; Vol. 276, No. 2. pp. 756-761.
@article{d4b81257d35a4a54b9fe94426113d3e0,
title = "Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo",
abstract = "Knock-out of the gene coding for caveolin-1, the main organizer of caveolae, has not yet been performed. We devised a strategy to knock-down caveolin-1 gene expression using antisense oligodeoxynucleotides (ODNs). Seven ODNs, covering different regions of caveolin-1 mRNA, were screened by Western blot analysis of caveolin-1 levels. The most active and specific was found to reduce caveolin-1 protein levels by 70{\%} at 1 μM concentration and its action, as demonstrated by a marked reduction (about 50{\%}) in caveolin-1 mRNA levels, was due to a true antisense mechanism. In HUVEC treated with the active ODN, caveolae were undetectable by confocol and electron microscopy, while their number was not affected when cells were treated with a scrambled ODN. Using the fibrin gel 3 D angiogenesis test we established that the active (but not the scrambled) ODN strongly suppressed capillary-like tube formation. Moreover, an antisense tailored against chicken caveolin-1 mRNA, when tested using the chorio-allantoic membrane technique, dramatically reduced vessel formation at doses (10-20 μg) under which control ODNs were ineffective and devoid of toxicity. Thus, it is likely that caveolin-1 down regulation, followed by caveolae disruption, impairs angiogenesis in vitro and in vivo. (C) 2000 Academic Press.",
keywords = "Angiogenesis, Antisense oligonucleotides, Caveolin",
author = "Cristiana Griffoni and Enzo Spisni and Spartaco Santi and Massimo Riccio and Tiziana Guarnieri and Vittorio Tomasi",
year = "2000",
month = "9",
day = "24",
doi = "10.1006/bbrc.2000.3484",
language = "English",
volume = "276",
pages = "756--761",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Knockdown of caveolin-1 by antisense oligonucleotides impairs angiogenesis in vitro and in vivo

AU - Griffoni, Cristiana

AU - Spisni, Enzo

AU - Santi, Spartaco

AU - Riccio, Massimo

AU - Guarnieri, Tiziana

AU - Tomasi, Vittorio

PY - 2000/9/24

Y1 - 2000/9/24

N2 - Knock-out of the gene coding for caveolin-1, the main organizer of caveolae, has not yet been performed. We devised a strategy to knock-down caveolin-1 gene expression using antisense oligodeoxynucleotides (ODNs). Seven ODNs, covering different regions of caveolin-1 mRNA, were screened by Western blot analysis of caveolin-1 levels. The most active and specific was found to reduce caveolin-1 protein levels by 70% at 1 μM concentration and its action, as demonstrated by a marked reduction (about 50%) in caveolin-1 mRNA levels, was due to a true antisense mechanism. In HUVEC treated with the active ODN, caveolae were undetectable by confocol and electron microscopy, while their number was not affected when cells were treated with a scrambled ODN. Using the fibrin gel 3 D angiogenesis test we established that the active (but not the scrambled) ODN strongly suppressed capillary-like tube formation. Moreover, an antisense tailored against chicken caveolin-1 mRNA, when tested using the chorio-allantoic membrane technique, dramatically reduced vessel formation at doses (10-20 μg) under which control ODNs were ineffective and devoid of toxicity. Thus, it is likely that caveolin-1 down regulation, followed by caveolae disruption, impairs angiogenesis in vitro and in vivo. (C) 2000 Academic Press.

AB - Knock-out of the gene coding for caveolin-1, the main organizer of caveolae, has not yet been performed. We devised a strategy to knock-down caveolin-1 gene expression using antisense oligodeoxynucleotides (ODNs). Seven ODNs, covering different regions of caveolin-1 mRNA, were screened by Western blot analysis of caveolin-1 levels. The most active and specific was found to reduce caveolin-1 protein levels by 70% at 1 μM concentration and its action, as demonstrated by a marked reduction (about 50%) in caveolin-1 mRNA levels, was due to a true antisense mechanism. In HUVEC treated with the active ODN, caveolae were undetectable by confocol and electron microscopy, while their number was not affected when cells were treated with a scrambled ODN. Using the fibrin gel 3 D angiogenesis test we established that the active (but not the scrambled) ODN strongly suppressed capillary-like tube formation. Moreover, an antisense tailored against chicken caveolin-1 mRNA, when tested using the chorio-allantoic membrane technique, dramatically reduced vessel formation at doses (10-20 μg) under which control ODNs were ineffective and devoid of toxicity. Thus, it is likely that caveolin-1 down regulation, followed by caveolae disruption, impairs angiogenesis in vitro and in vivo. (C) 2000 Academic Press.

KW - Angiogenesis

KW - Antisense oligonucleotides

KW - Caveolin

UR - http://www.scopus.com/inward/record.url?scp=0034710841&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034710841&partnerID=8YFLogxK

U2 - 10.1006/bbrc.2000.3484

DO - 10.1006/bbrc.2000.3484

M3 - Article

C2 - 11027543

AN - SCOPUS:0034710841

VL - 276

SP - 756

EP - 761

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 2

ER -