Involvement of lysosomal degradation in VEGF-C-induced down-regulation of VEGFR-3

• Published in: FEBS letters 2014-11, Vol.588 (23), p.4357-4363
• Main Authors: Han, Kyu-Yeon, Chang, Jin-Hong, Dugas-Ford, Jennifer, Alexander, Jonathan S., Azar, Dimitri T.
• Format: Article
• Language: English
• Subjects: bovine serum albumin; BSA; c-Jun N-terminal kinase; Cell Line; DAPT; Down-Regulation - drug effects; endothelial cell; Endothelial Cells - cytology; Endothelial Cells - drug effects; Endothelial Cells - metabolism; ERK; extracellular signal-regulated kinase; Extracellular Signal-Regulated MAP Kinases - metabolism; Humans; JNK; JNK Mitogen-Activated Protein Kinases - metabolism; LEC; leupeptin, pepstatin, and E64; Ligands; LPE; lymphatic endothelial cell; Lysosomal degradation; Lysosomes - drug effects; Lysosomes - metabolism; matrix metalloproteinase; MMP; N-[N-(3,5-Difluorophenacetyl)-L-alanyl]-S-phenylglycine t-butyl ester; p38 Mitogen-Activated Protein Kinases - metabolism; PCR; Phosphorylation - drug effects; placenta growth factor; PlGF; polymerase chain reaction; Protein Kinase Inhibitors - pharmacology; Protein Transport - drug effects; Proteolysis - drug effects; receptor tyrosine kinase; RNA, Messenger - genetics; RNA, Messenger - metabolism; RTK; Signal Transduction - drug effects; simian virus 40; simian virus 40-immortalized lymphatic endothelial cell; SV-LEC; SV40; vascular endothelial growth factor; Vascular Endothelial Growth Factor C - metabolism; vascular endothelial growth factor receptor; Vascular Endothelial Growth Factor Receptor-3 - genetics; Vascular Endothelial Growth Factor Receptor-3 - metabolism; VEGF; VEGF receptor 3; VEGF-C; VEGFR
• ISSN: 0014-5793; 1873-3468
• DOI: 10.1016/j.febslet.2014.09.034

•VEGF-C specifically down-regulates VEGFR-3 mRNA and protein in lymphatic endothelial cells.•Down-regulation is mediated by VEGF-C binding to VEGFR-3 and VEGFR-3 auto-phosphorylation.•Down-regulation does not require MMP activity or Notch signaling.•Lysosomal protease inhibitors decrease the down-regulatory effect on VEGFR-3 protein levels.•Results indicate a ligand-induced receptor down-regulation mechanism via lysosomal activity. The vascular endothelial growth factor (VEGF)-C-induced down-regulation of VEGF receptor (VEGFR)-3 is important in lymphangiogenesis. Here, we demonstrate that VEGF-C, -D, and -C156S, but not VEGF-A, down-regulate VEGFR-3. VEGF-C stimulates VEGFR-3 tyrosyl phosphorylation and transient phosphorylation of extracellular signal-regulated kinase (ERK), p38, and c-Jun N-terminal kinases in lymphatic endothelial cells. VEGF-C-induced down-regulation of VEGFR-3 was blocked by a VEGF-C trap, tyrosine kinase inhibitor, and leupeptin, pepstatin, and E64 (LPE), but was unaffected by Notch 1 activator and γ-secretase inhibitors. Our findings indicate that VEGF-C down-regulates VEGFR-3 in lymphatic endothelial cells through VEGFR-3 kinase activation and, in part, via lysosomal degradation.