title: Expression of tissue-type plasminogen activator and its inhibitor couples with development of capillary network by human microvascular endothelial cells on Matrigel.
abstract: Human omental microvascular endothelial (HOME) cells seeded on Matrigel begin to migrate within 1 h, forming honeycomb-like structures and capillary-like networks within 18 h. Cross-sections of the capillary networks show them to be tube-like structures. Northern blot analysis showed that tissue-type plasminogen activator (t-PA) mRNA synthesis increased from the initial state at 0 h after seeding on Matrigel, reaching a steady state after 4 h. This elevated cellular t-PA mRNA level decreased markedly at 24 h. In contrast, the cellular plasminogen activator inhibitor-1 (PAI-1) mRNA level demonstrated biphasic curves during the 24 h after seeding on Matrigel: the PAI-1 mRNA level was increased eightfold initially at 4 h over that at 0 h, then declined, and again secondarily increased to greater than tenfold at 18 h. Cellular levels of both 72 kD type IV collagenase and tissue inhibitor of metalloproteinase (TIMP-2) mRNA were increased only a slightly within 2-4 h. These elevated mRNA levels were maintained for 18 h, while the TIMP-1 mRNA level increased up to 18 h, reaching around three times the level at 0 h. However, on collagen-coated dishes, cellular levels of t-PA, PAI-1, 72 kD type IV collagenase, TIMP-1, and TIMP-2 mRNA were not greatly changed during incubation for 24 h. On Matrigel, the cellular t-PA mRNA level at 18 h after seeding was greatly increased when treated with specific anti-transforming growth factor-beta (TGF-beta) antibody. In contrast, both PAI-1 and TIMP-1 mRNA levels at 18 h were reduced in the presence of anti-TGF-beta antibody. Development of the capillary network on Matrigel was inhibited in the presence of anti-t-PA antibody. Epidermal growth factor (EGF) enhanced t-PA gene expression and TGF-beta inhibited its expression in HOME cells cultured on collagen-coated dishes. On the other hand, TGF-beta enhanced cellular expression of the PAI-1 gene. The formation of a capillary network by HOME cells on Matrigel appears to be balanced by angiogenic EGF and anti-angiogenic TGF-beta through modulation of PA activity.
authors: Department of Biochemistry, Kyushu University School of Medicine, Fukuoka, Japan.
authors: K
authors: K
authors: Ito
authors: M
authors: M
authors: Ryuto
authors: S
authors: S
authors: Ushiro
authors: M
authors: M
authors: Ono
authors: A
authors: A
authors: Sugenoya
authors: A
authors: A
authors: Kuraoka
authors: Y
authors: Y
authors: Shibata
authors: M
authors: M
authors: Kuwano
mesh_terms: D002196:Capillaries / Q000254:growth & development* / Q000378:metabolism; D003094:Collagen; D017364:Collagenases / Q000096:biosynthesis; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology / Q000254:growth & development* / Q000378:metabolism; D004815:Epidermal Growth Factor / Q000494:pharmacology; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D006023:Glycoproteins / Q000096:biosynthesis; D006801:Humans; D007700:Kinetics; D007797:Laminin; D020780:Matrix Metalloproteinase 9; D061965:Matrix Metalloproteinase Inhibitors; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D010537:Peritoneum / Q000166:cytology; D017395:Plasminogen Activator Inhibitor 1 / Q000096:biosynthesis*; D014176:Protein Biosynthesis; D011509:Proteoglycans; D012333:RNA, Messenger / Q000096:biosynthesis; D019716:Tissue Inhibitor of Metalloproteinase-2; D019714:Tissue Inhibitor of Metalloproteinases; D010959:Tissue Plasminogen Activator / Q000096:biosynthesis* / Q000494:pharmacology; D016212:Transforming Growth Factor beta / Q000502:physiology
chemical_list: D004338:Drug Combinations; D006023:Glycoproteins; D007797:Laminin; D061965:Matrix Metalloproteinase Inhibitors; D017395:Plasminogen Activator Inhibitor 1; D011509:Proteoglycans; D012333:RNA, Messenger; D019714:Tissue Inhibitor of Metalloproteinases; D016212:Transforming Growth Factor beta; C059485:matrigel; D019716:Tissue Inhibitor of Metalloproteinase-2; D004815:Epidermal Growth Factor; D003094:Collagen; D010959:Tissue Plasminogen Activator; D017364:Collagenases; D008666:Metalloendopeptidases; D020780:Matrix Metalloproteinase 9