Vascular Endothelial Growth Factor-D Activates VEGFR-3 Expressed in Osteoblasts Inducing Their Differentiation

In vivo imaging of lymphatic vessels in development, wound healing, inflammation, and tumor metastasis

“…Although Vegfr3 is also expressed, to some extent, in the tip cells of the newly formed blood capillaries (21)(22)(23) and in fenestrated endothelium (24), it is still one of the best markers for all lymphatic endothelial cells. Outside the vascular system Vegfr3 expression has been described in subpopulations of monocytes and macrophages (25) and in embryonic osteoblasts and neural progenitors (26,27). We have validated this reporter for tracking lymphatic vessels in vivo during development, and lymphangiogenesis after administration of specific growth factors and inhibitors, in wound healing, inflammation and tumor dissemination.…”

Downregulation of VEGF-C expression in lung and colon cancer cells decelerates tumor growth and inhibits metastasis via multiple mechanisms

“…It binds to Flt4/VEGF receptor 3 (VEGFR3), a receptor tyrosine kinase that is expressed in lymphatic endothelial cells (Joukov et al, 1996), certain non-endothelial cells such as osteoblasts (Orlandini et al, 2006) and many cancer cell types (Strizzi et al, 2001;Jackson et al, 2002;Su et al, 2006;Timoshenko et al, 2007). With significantly lower affinity VEGF-C interacts with VEGFR2, which is primarily involved in the induction of hemangiogenesis (Skobe et al, 1999;Shibuya and Claesson-Welsh, 2006) and coreceptors, including the neuropilin-2 expressed on veins and lymphatic vessels (Karkkainen et al, 2001;Karpanen et al, 2006;Xu et al, 2010).…”

Morphogenesis of human endothelial cells is inhibited by DAB2 via Src

“…Cells were seeded on glass coverslips and fixed in 3% paraformaldehyde. Specimens were treated as previously described [25]. The primary antibodies used were: rabbit anti-von Willebrand factor (Dako), rat anti-CD31 (PharMingen), mouse anti-Dab2/p96 (Transduction Laboratories), rabbit anti-phospho-Y925 FAK, rabbit anti-Src (32G6), and rabbit anti-phospho-Y416 Src (Cell Signaling).…”

“…Small hairpin (shRNA) cassette was cloned, and the recombinant lentiviruses were produced as previously described [25]. Briefly, oligonucleotides coding for human DAB2 (GeneBank accession number NM_001343) and unrelated (synthetic construct, GeneBank accession Abbreviations: DAB2, Disabled-2; HUVEC, human umbilical vein endothelial cells; MAPK, mitogen-activated protein kinase; ERK, extracellular-signal regulated kinase; FAK, focal adhesion kinase; JNK, c-Jun NH 2 -terminal kinase number DQ092361) shRNA were designed to contain a sense strand 5 0 -GTCCATACAGAATGGCGTAAA-3 0 (DAB2, clone 22), 5 0 -GTATCTGAAAGAGAACAGAAC-3 0 (DAB2, clone 47), and 5 0 -GCCACAAGTTCAGCGTGTC-3 0 (unrelated) followed by a spacer, their reverse complementary strand, and an RNA polymerase III transcriptional stop signal.…”