Angiogenesis

, 8:315 | Cite as

Src, Fyn and Yes play differential roles in VEGF-mediated endothelial cell events

Article

Abstract

Widely coexpressed Src family kinase (SFK) members Src, Fyn and Yes are involved in various cellular events, often acting downstream of receptor tyrosine kinases, such as vascular endothelial growth factor (VEGF) receptors. They are well known for their functional redundancy; any unique features remain largely undefined. Utilizing RNA interference, we have selectively knocked down Src, Fyn and Yes in human retinal microvascular endothelial cells (HRMECs). Cells with single SFK knockdown showed that all three kinases were required for VEGF mitogenic signaling. VEGF-induced cell migration was significantly increased in Fyn-deficient cells and decreased in Yes-deficient cells. Selective interference of Fyn, but not Src or Yes, impaired VEGF-induced tube formation in HRMECs. Cells in which all three SFKs were targeted showed significant inhibition of all three cellular events. In addition, interference of Src, Fyn and Yes did not affect the anti-apoptotic effect of VEGF in HRMECs, as determined by DNA fragmentation analysis. These results provide direct evidence that Src, Fyn and Yes maintain distinct properties in the regulation of VEGF-mediated endothelial cell events.

Keywords

Fyn RNA interference Src vascular endothelial growth factor Yes 

Notes

Acknowledgements

We thank Dr Abboud J. Ghalayini and Dr Jin Chen for insightful discussion and suggestion. We thank Dr Gary W. McCollum for experimental support. We thank Dr Derya Unutmaz for sharing the NucleofectorTM Device, and Dr Robbert J. Slebos for sharing the SynergyTM HT multi-detection microplate reader. We thank the Vanderbilt University Medical Center Cell Imaging Shared Resource for training and sharing of microscopes and software.

This work was supported by NIH grants EY07533 and EY08126 and a grant from Research to Prevent Blindness, Inc.

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Copyright information

© Springer Science+Business Media, Inc. 2005

Authors and Affiliations

  1. 1.Department of Ophthalmology and Visual Sciences, Department of Cell and Developmental BiologyVanderbilt University School of MedicineNashvilleUSA
  2. 2.Department of Ophthalmology and Visual SciencesVanderbilt University School of MedicineNashvilleUSA

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