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Protein kinase C promotes angiogenic activity of human endothelial cells via induction of vascular endothelial growth factor
- Source :
- Cardiovascular Research. 78:349-355
- Publication Year :
- 2008
- Publisher :
- Oxford University Press (OUP), 2008.
-
Abstract
- Aims Protein kinase C (PKC) plays an important role in the regulation of angiogenesis. However, downstream targets of PKC in endothelial cells are poorly defined. Methods and results mRNA expression of vascular endothelial growth factor (VEGF) was analysed by quantitative real-time RT-PCR in human umbilical vein endothelial cells (HUVEC) and HUVEC-derived EA.hy 926 cells. siRNA was used to knockdown PKC isoforms and VEGF. Matrigel tube formation assay was used to analyse the angiogenic activity of endothelial cells. Phorbol-12-myristate-13-acetate (PMA) enhanced the ability of HUVEC to organize into tubular networks when plated on Matrigel, a phenomenon that could be prevented by PKC inhibitors. PMA markedly increased the expression of VEGF in HUVEC and EA.hy 926 cells. The enhancement in VEGF expression was prevented by PKC inhibitors and by an inhibitor of the Erk1/2 pathway. PMA-induced tube formation was reduced by inhibition of the VEGF receptor kinase, or by VEGF knockdown. PMA led to an activation of PKC isoforms α, δ and e in HUVEC. Knockdown of PKC α diminished PMA-induced VEGF expression and angiogenesis. Also endothelial progenitor cells isolated from human peripheral blood showed enhanced VEGF expression and improved angiogenic activity in response to PKC activation. Moreover, incubation of HUVEC with VEGF led to PKC α activation and PKC-dependent VEGF upregulation. Conclusions PKC α activation promotes angiogenic activity of human endothelial cells. This is likely to be largely mediated by induction of VEGF. VEGF enhances its own expression via a PKC α-dependent positive feedback mechanism.
- Subjects :
- Vascular Endothelial Growth Factor A
medicine.medical_specialty
Protein Kinase C-alpha
Time Factors
Physiology
Angiogenesis
medicine.medical_treatment
Blotting, Western
Carbazoles
Neovascularization, Physiologic
Biology
Polymerase Chain Reaction
Cell Line
chemistry.chemical_compound
Physiology (medical)
Internal medicine
medicine
Humans
RNA, Messenger
RNA, Small Interfering
Cell Shape
Protein Kinase Inhibitors
Cells, Cultured
Protein kinase C
Tube formation
Matrigel
Stem Cells
Growth factor
Endothelial Cells
Up-Regulation
Cell biology
Enzyme Activation
Endothelial stem cell
Vascular endothelial growth factor
Autocrine Communication
Vascular endothelial growth factor A
Receptors, Vascular Endothelial Growth Factor
Endocrinology
chemistry
Tetradecanoylphorbol Acetate
Angiogenesis Inducing Agents
Fibroblast Growth Factor 2
RNA Interference
Cardiology and Cardiovascular Medicine
Subjects
Details
- ISSN :
- 00086363
- Volume :
- 78
- Database :
- OpenAIRE
- Journal :
- Cardiovascular Research
- Accession number :
- edsair.doi.dedup.....e9c10739c6f0c47b900eefff0231b0e4