Isolated lymphatic endothelial cells transduce growth, survival and migratory signals via the VEGF-C/D receptor VEGFR-3.
Vascular endothelial growth factor receptor-3 (VEGFR-3/Flt4) binds two known members of the VEGF ligand family, VEGF-C and VEGF-D, and has a critical function in the remodelling of the primary capillary vasculature of midgestation embryos. Later during development, VEGFR-3 regulates the growth and maintenance of the lymphatic vessels. In the present study, we have isolated and cultured stable lineages of blood vascular and lymphatic endothelial cells from human primary microvascular endothelium by using antibodies against the extracellular domain of VEGFR-3. We show that VEGFR-3 stimulation alone protects the lymphatic endothelial cells from serum deprivation-induced apoptosis and induces their growth and migration. At least some of these signals are transduced via a protein kinase C-dependent activation of the p42/p44 MAPK signalling cascade and via a wortmannin-sensitive induction of Akt phosphorylation. These results define the critical role of VEGF-C/VEGFR-3 signalling in the growth and survival of lymphatic endothelial cells. The culture of isolated lymphatic endothelial cells should now allow further studies of the molecular properties of these cells.
Mäkinen
T
T
Molecular/Cancer Biology Laboratory and Ludwig Institute for Cancer Research, and Helsinki University Hospital, Biomedicum Helsinki, University of Helsinki, FIN-00014 Helsinki, Finland.
Veikkola
T
T
Mustjoki
S
S
Karpanen
T
T
Catimel
B
B
Nice
E C
EC
Wise
L
L
Mercer
A
A
Kowalski
H
H
Kerjaschki
D
D
Stacker
S A
SA
Achen
M G
MG
Alitalo
K
K
D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D015374:Biosensing Techniques; D002196:Capillaries / Q000196:embryology / Q000502:physiology; D002455:Cell Division; D002465:Cell Movement; D002470:Cell Survival; D002478:Cells, Cultured; D016228:Endothelial Growth Factors / Q000494:pharmacology* / Q000502:physiology*; D004727:Endothelium / Q000166:cytology / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000502:physiology*; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D008208:Lymphatic System / Q000166:cytology / Q000502:physiology*; D020935:MAP Kinase Signaling System / Q000502:physiology*; D008833:Microcirculation / Q000502:physiology; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics; D020794:Receptor Protein-Tyrosine Kinases / Q000235:genetics / Q000502:physiology*; D017978:Receptors, Growth Factor / Q000235:genetics / Q000502:physiology*; D040262:Receptors, Vascular Endothelial Growth Factor; D012867:Skin / Q000098:blood supply; D014471:Umbilical Veins; D042582:Vascular Endothelial Growth Factor C; D042643:Vascular Endothelial Growth Factor D; D040321:Vascular Endothelial Growth Factor Receptor-3
D016228:Endothelial Growth Factors; D012333:RNA, Messenger; D017978:Receptors, Growth Factor; D042582:Vascular Endothelial Growth Factor C; D042643:Vascular Endothelial Growth Factor D; D020794:Receptor Protein-Tyrosine Kinases; D040262:Receptors, Vascular Endothelial Growth Factor; D040321:Vascular Endothelial Growth Factor Receptor-3; D011493:Protein Kinase C; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D020928:Mitogen-Activated Protein Kinases