Ets-1 regulates angiogenesis by inducing the expression of urokinase-type plasminogen activator and matrix metalloproteinase-1 and the migration of vascular endothelial cells.
The coordinate induction of protease activities and cell migration is a principal feature of endothelial cells (ECs) invading the interstitial space in the initial step of angiogenesis. However, the molecular mechanisms of these events are not fully characterized. Ets-1 is a member of the ets gene family of transcription factors, which binds to the Ets binding motif in the cis-acting elements and regulates the expression of certain genes. Four typical angiogenic growth factors, aFGF, bFGF, VEGF, and EGF, induced the expression of ets-1 mRNA in either human umbilical vein endothelial cells (HUVECs), ECV-304 cells (immortalized HUVECs), or human omental microvascular endothelial cells (HOMECs). The expression of ets-1 reached its maximum at 2 hr after factor addition and then decreased to the basal level by 12 hr. For characterization of the role of Ets-1 in angiogenesis, ets-1 antisense and sense oligodeoxynucleotides (ODNs) were constructed. The ets-1 antisense ODN but not sense ODN efficiently blocked the synthesis of Ets-1 protein by human ECs in response to angiogenic growth factors. Moreover, the ets-1 antisense ODN but not sense ODN almost completely abolished the binding of endothelial cell extract to DNA containing the Ets binding motif. The expression of urokinase-type plasminogen activator and matrix metalloproteinase-1 and the migration of ECs in response to growth factors were significantly inhibited by ets-1 antisense ODN but not by sense ODN. Tube formation by HOMECs in type 1 collagen gel stimulated with EGF was abrogated by ets-1 antisense ODN. Finally, the expression of Ets-1 protein in ECs during angiogenesis in vivo was confirmed by an immunohistochemical analysis using a murine angiogenesis model. These results indicate that the induction of ets-1 mRNA is a mutual phenomenon in ECs stimulated with angiogenic growth factors. Ets-1 appears to play an important role in angiogenesis, regulating the expression of proteases and the migration of ECs.
Iwasaka
C
C
Department of Vascular Biology, Tohoku University, Sendai, Japan.
Tanaka
K
K
Abe
M
M
Sato
Y
Y
D000818:Animals; D001808:Blood Vessels / Q000166:cytology*; D002465:Cell Movement; D002478:Cells, Cultured; D003094:Collagen; D017364:Collagenases / Q000378:metabolism*; D004268:DNA-Binding Proteins / Q000378:metabolism; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology*; D004790:Enzyme Induction; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005346:Fibroblast Growth Factors / Q000494:pharmacology; D015870:Gene Expression; D006801:Humans; D007797:Laminin; D020781:Matrix Metalloproteinase 1; D051379:Mice; D016376:Oligonucleotides, Antisense / Q000494:pharmacology; D011509:Proteoglycans; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins / Q000502:physiology*; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger / Q000235:genetics; D014157:Transcription Factors / Q000502:physiology*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D004268:DNA-Binding Proteins; D004338:Drug Combinations; C494041:ETS1 protein, human; C494042:Ets1 protein, mouse; D007797:Laminin; D016376:Oligonucleotides, Antisense; D011509:Proteoglycans; D050784:Proto-Oncogene Protein c-ets-1; D011518:Proto-Oncogene Proteins; D050783:Proto-Oncogene Proteins c-ets; D012333:RNA, Messenger; D014157:Transcription Factors; C059485:matrigel; D005346:Fibroblast Growth Factors; D004815:Epidermal Growth Factor; D003094:Collagen; D014568:Urokinase-Type Plasminogen Activator; D017364:Collagenases; D020781:Matrix Metalloproteinase 1