Role of JNK in network formation of human lung microvascular endothelial cells.
The signaling mechanisms in vasculogenesis and/or angiogenesis remain poorly understood, limiting the ability to regulate growth of new blood vessels in vitro and in vivo. Cultured human lung microvascular endothelial cells align into tubular networks in the three-dimensional matrix, Matrigel. Overexpression of MAPK phosphatase-1 (MKP-1), an enzyme that inactivates the ERK, JNK, and p38 pathways, inhibited network formation of these cells. Adenoviral-mediated overexpression of recombinant MKP-3 (a dual specificity phosphatase that specifically inactivates the ERK pathway) and dominant negative or constitutively active MEK did not attenuate network formation in Matrigel compared with negative controls. This result suggested that the ERK pathway may not be essential for tube assembly, a conclusion which was supported by the action of specific MEK inhibitor PD 184352, which also did not alter network formation. Inhibition of the JNK pathway using SP-600125 or l-stereoisomer (l-JNKI-1) blocked network formation, whereas the p38 MAPK blocker SB-203580 slightly enhanced it. Inhibition of JNK also attenuated the number of small vessel branches in the developing chick chorioallantoic membrane. Our results demonstrate a specific role for the JNK pathway in network formation of human lung endothelial cells in vitro while confirming that it is essential for the formation of new vessels in vivo.
Medhora
Meetha
M
Division of Pulmonary and Critical Care, Department of Medicine, Medical College of Wisconsin, Milwaukee, WI 53226, USA. medhoram@mcw.edu
Dhanasekaran
Anuradha
A
Pratt
Phillip F
PF
Cook
Craig R
CR
Dunn
Laurel K
LK
Gruenloh
Stephanie K
SK
Jacobs
Elizabeth R
ER
D000818:Animals; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D003094:Collagen; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology / Q000201:enzymology*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005419:Flavonoids / Q000494:pharmacology; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007797:Laminin; D048670:MAP Kinase Kinase 4 / Q000378:metabolism*; D008833:Microcirculation; D018919:Neovascularization, Physiologic; D011509:Proteoglycans; D011652:Pulmonary Circulation / Q000502:physiology*
D004338:Drug Combinations; D004791:Enzyme Inhibitors; D005419:Flavonoids; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D049452:Green Fluorescent Proteins; D003094:Collagen; D048670:MAP Kinase Kinase 4; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one