Effect of vascular endothelial growth factor and epidermal growth factor on iatrogenic apoptosis in human endothelial cells

Maria Cristina Vinci, Barbara Visentin, Federico Cusinato, Giovanni Battista Nardelli, Lucia Trevisi, Sisto Luciani

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

To study the effect of growth factors on iatrogenic apoptosis, we examined the influence of vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) on staurosporine-induced apoptosis in primary cultures of human umbilical vein endothelial cells (HUVEC). Apoptosis was evaluated by a cell viability test, the TUNEL-POD assay and the activation of the pro-apoptotic caspase-3. Staurosporine (10-100nM) caused the activation of caspase-3. This effect was manifest after 2hr of incubation and reached its maximum after 5hr. Severe loss of viability followed within 18hr. VEGF or EGF (10-100ng/mL) added together with staurosporine decreased the activation of caspase-3. The loss of viability was 24hr delayed. The action of growth factors was observed at 1% serum concentration but also at concentration optimal for HUVEC survival (10%, v/v). Furthermore, the inhibition of PI-3 kinase (PI-3K) by wortmannin or LY294002 as well as the inhibition of MEK by PD098059 or U0126 prevented the protective effect of VEGF and EGF. Western blotting analysis showed that after 3hr of incubation with staurosporine the level of the anti-apoptotic protein Mcl-1 decreased and this effect was reverted by VEGF. It is concluded that VEGF and EGF antagonize the pro-apoptotic action of staurosporine by the combined signalling of PI-3K and ERKs pathways.

Original languageEnglish
Pages (from-to)277-284
Number of pages8
JournalBiochemical Pharmacology
Volume67
Issue number2
DOIs
Publication statusPublished - Jan 15 2004

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Staurosporine
Endothelial cells
Epidermal Growth Factor
Vascular Endothelial Growth Factor A
Endothelial Cells
Apoptosis
Caspase 3
Chemical activation
Human Umbilical Vein Endothelial Cells
Phosphatidylinositol 3-Kinases
Cell Survival
Intercellular Signaling Peptides and Proteins
Apoptosis Regulatory Proteins
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
MAP Kinase Signaling System
Mitogen-Activated Protein Kinase Kinases
In Situ Nick-End Labeling
Cell culture
Assays
Western Blotting

Keywords

  • Apoptosis
  • EGF
  • ERKs
  • PI-3 kinase
  • Staurosporine
  • VEGF

ASJC Scopus subject areas

  • Pharmacology

Cite this

Effect of vascular endothelial growth factor and epidermal growth factor on iatrogenic apoptosis in human endothelial cells. / Vinci, Maria Cristina; Visentin, Barbara; Cusinato, Federico; Nardelli, Giovanni Battista; Trevisi, Lucia; Luciani, Sisto.

In: Biochemical Pharmacology, Vol. 67, No. 2, 15.01.2004, p. 277-284.

Research output: Contribution to journalArticle

Vinci, Maria Cristina ; Visentin, Barbara ; Cusinato, Federico ; Nardelli, Giovanni Battista ; Trevisi, Lucia ; Luciani, Sisto. / Effect of vascular endothelial growth factor and epidermal growth factor on iatrogenic apoptosis in human endothelial cells. In: Biochemical Pharmacology. 2004 ; Vol. 67, No. 2. pp. 277-284.
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