Angiostatin effects on endothelial cells mediated by ceramide and RhoA.
Angiostatin is a cleavage product of plasminogen that has anti-angiogenic properties. We investigated whether the effects of angiostatin on endothelial cells are mediated by ceramide, a lipid implicated in endothelial cell signaling. Our results demonstrate that angiostatin produces a transient increase in ceramide that correlates with actin stress fiber reorganization, detachment and death. DNA array expression analysis performed on ceramide-treated human endothelial cells demonstrated induction of certain genes involved in cytoskeleton organization. Specifically, we report that treatment with angiostatin or ceramide results in the activation of RhoA, an important effector of cytoskeletal structure. We also show that treatment of endothelial cells with the antioxidant N-acetylcysteine abrogates morphological changes and cytotoxic effects of treatment with angiostatin or ceramide. These findings support a model in which angiostatin induces a transient rise in ceramide, RhoA activation and free radical production.
Gupta
N
N
Department of Surgery, University of Chicago, MC 9006, 5758 South Maryland Avenue, Chicago, IL 60637, USA.
Nodzenski
E
E
Khodarev
N N
NN
Yu
J
J
Khorasani
L
L
Beckett
M A
MA
Kufe
D W
DW
Weichselbaum
R R
RR
D043168:Angiostatins; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D003599:Cytoskeleton / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004730:Endothelium, Vascular / Q000166:cytology*; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D015151:Immunoblotting; D007700:Kinetics; D060850:LIM-Homeodomain Proteins; D008858:Microscopy, Phase-Contrast; D009419:Nerve Tissue Proteins / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010446:Peptide Fragments / Q000502:physiology*; D010958:Plasminogen / Q000502:physiology*; D021381:Protein Transport; D015398:Signal Transduction; D013110:Sphingosine / Q000031:analogs & derivatives / Q000378:metabolism*; D013997:Time Factors; D014157:Transcription Factors; D014471:Umbilical Veins / Q000166:cytology; D020742:rhoA GTP-Binding Protein / Q000378:metabolism*
D018398:Homeodomain Proteins; C491186:LHX5 protein, human; D060850:LIM-Homeodomain Proteins; C064769:N-acetylsphingosine; D009419:Nerve Tissue Proteins; D010446:Peptide Fragments; D014157:Transcription Factors; D043168:Angiostatins; D010958:Plasminogen; D020742:rhoA GTP-Binding Protein; D013110:Sphingosine