Skip to Content
Merck
CN
  • Activin receptor-like kinase 1 inhibits human microvascular endothelial cell migration: potential roles for JNK and ERK.

Activin receptor-like kinase 1 inhibits human microvascular endothelial cell migration: potential roles for JNK and ERK.

Journal of cellular physiology (2007-07-11)
Laurent David, Christine Mallet, Bruno Vailhé, Samy Lamouille, Jean-Jacques Feige, Sabine Bailly
ABSTRACT

Activin receptor-like kinase 1 (ALK1) is an endothelial-specific type I receptor of the TGFbeta receptor family that is implicated in angiogenesis and in the pathogenesis of the vascular disease, hereditary hemorrhagic telangiectasia (HHT). In the absence of a specific ligand, ALK1 cellular functions have been mainly studied through the use of a constitutively active form of this receptor (ALK1ca) and are still debated. We previously reported that ALK1ca inhibits proliferation and migration of human endothelial cells suggesting that ALK1 plays an important role in the maturation phase of angiogenesis (Lamouille et al., 2002, Blood 100: 4495-4501). In the present work, we further analyzed the role of ALK1 in the migration of human dermal microvascular endothelial cell (HMVEC-d) and observed that silencing endogenous ALK1 expression with siRNAs accelerates endothelial cell migration in the wound assay. Further, we demonstrate that ALK1-induced inhibition of migration is Smad-independent. Using a panel of kinase inhibitors, we found that HMVEC-d wound closure was completely inhibited by a JNK inhibitor and to a lower degree by an ERK kinase inhibitor. Further, HMVEC-d wounding induced activation of both JNK and ERK, and these were inhibited by ALK1ca expression. Taken together, these results support a significant role for ALK1 as a negative regulator of endothelial cell migration and suggest the implication of JNK and ERK as mediators of this effect.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Y-27632-CAS 331752-47-7-Calbiochem, Y-27632A, CAS 331752-47-7, is a cell-permeable, reversible, inhibitor of Rho kinases (Ki = 140 nM for p160ROCK). Enhances survival & cloning efficiency of ESC without affecting their pluripotency.
Sigma-Aldrich
Wortmannin, Wortmannin, CAS 19545-26-7, is a cell-permeable, potent, selective, and irreversible inhibitor of PI3-Kinase (IC50 = 5 nM). Does not affect any upstream signaling events.
Millipore
SB 202190, SB 202190, CAS 152121-30-7, is a potent, reversible, competitive inhibitor of p38. Inhibits p38 phosphorylation of myelin basic protein. Blocks the activity of p38β (Ki = 16 nM; IC50 = 350 nM).
Sigma-Aldrich
LY 294002, LY294002, CAS 154447-36-6, is a cell-permeable, potent, reversible, and specific inhibitor of PI 3-kinase ((IC50 = 1.4 µM). Acts on the ATP-binding site.
Sigma-Aldrich
Anti-MAP Kinase (ERK-1, ERK-2) antibody produced in rabbit, whole antiserum
Sigma-Aldrich
PD 98059, PD 98059, CAS 167869-21-8, is a cell-permeable, selective & reversible inhibitor of MAP Kinase Kinase (MEK). Inhibits MAP Kinase activation and subsequent phosphorylation of MAP Kinase substrates.
Sigma-Aldrich
JNK Inhibitor II, JNK Inhibitor II. SP600125, CAS 129-56-6, is a potent, cell-permeable, selective, and ATP competitive inhibitor of c-Jun N-terminal kinase (JNK; IC50 = 40 nM for JNK-1 & JNK-2 & 90 nM for JNK-3).