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

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

Keywords

  • Angiogenesis
  • Antisense oligonucleotides
  • Caveolin

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

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