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VEGF masks BNIP3-mediated apoptosis of hypoxic endothelial cells

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Abstract

Hypoxia results in the apoptotic death of myocytes, neurons, and epithelial cells, through the actions of Bcl-2 and Nineteen kilodalton Interacting Protein-3 (BNIP3). On the contrary, endothelial cells are especially adept at surviving conditions of oxygen deprivation via up-regulation of vascular endothelial growth factor (VEGF) the most potent endothelial survival factor. Both VEGF and BNIP3 expression are transcriptionally regulated by hypoxia inducible factor and may antagonize each other’s affects in endothelial cells (ECs). Since factors that promote and inhibit apoptosis may be expressed at the same time in endothelial cells under hypoxic conditions, we decided to investigate whether VEGF and BNIP3 have opposing actions in endothelial cells. Human microvascular endothelial cells were exposed to hypoxic conditions in a Billups-Rothenburg chamber. Under hypoxic conditions BNIP3 expression by endothelial cells increased as measured by real-time PCR and immunoblot. After 48 h of hypoxia, EC apoptosis was assessed by flow cytometry and was lower than in corresponding normoxia serum starved controls. The increase in EC survival under hypoxic conditions corresponded with an increase in the expression of VEGF. Under normoxic conditions adenoviral BNIP3 over-expression promoted apoptosis of ECs; however, recombinant VEGF (100 pg/ml) antagonized the BNIP3 apoptosis promoting affects. SiRNA knockdown of VEGF expression by hypoxic ECs resulted in increased apoptosis with a concomitant increase in BNIP3 expression. SiRNA knockdown of BNIP3 expression by hypoxic ECs reduced the increase in EC apoptosis as a result of VEGF knockdown. We conclude that under hypoxic conditions VEGF counteracts and masks the apoptosis promoting affects of BNIP3.

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Acknowledgments

P. J. was supported by a fellowship from the Heart and Stroke Foundation of Canada. This work was supported by grants from the Canadian Institutes of Health Research [MOP-42402 to L.A.K., MOP-57726 to D.J.S.]; and the Heart and Stroke Foundation of Alberta, Nunavut, & NWT [grant-in aid to P. J.].

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Authors and Affiliations

  1. St. Michael’s Hospital, Toronto, ON, Canada

    Paul Jurasz & Duncan J. Stewart

  2. University of Toronto, Toronto, ON, Canada

    Duncan J. Stewart

  3. McLaughlin Centre for Molecular Medicine, Toronto, ON, Canada

    Duncan J. Stewart

  4. Ottawa Health Research Institute, The Ottawa Hospital, University of Ottawa, Ottawa, ON, Canada

    Duncan J. Stewart

  5. Department of Pharmacology and Therapeutics, Institute of Cardiovascular Sciences, St. Boniface General Hospital Research Centre, University of Manitoba, Winnipeg, MN, Canada

    Natasha Yurkova & Lorrie Kirshenbaum

  6. Faculty of Pharmacy and Pharmaceutical Sciences, 6152 Dentistry/Pharmacy Centre, University of Alberta, Edmonton, AB, T6G 2N8, Canada

    Paul Jurasz

  7. Department of Pharmacology, Faculty of Medicine and Dentistry, University of Alberta, Edmonton, AB, T6G 2N8, Canada

    Paul Jurasz

  8. Cardiovascular Research Centre, University of Alberta, Edmonton, AB, Canada

    Paul Jurasz

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  1. Paul Jurasz
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  2. Natasha Yurkova
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  3. Lorrie Kirshenbaum
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  4. Duncan J. Stewart
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Correspondence to Paul Jurasz.

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Jurasz, P., Yurkova, N., Kirshenbaum, L. et al. VEGF masks BNIP3-mediated apoptosis of hypoxic endothelial cells. Angiogenesis 14, 199–207 (2011). https://doi.org/10.1007/s10456-011-9204-6

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  • DOI: https://doi.org/10.1007/s10456-011-9204-6

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