Expression of tissue-type plasminogen activator and its inhibitor couples with development of capillary network by human microvascular endothelial cells on Matrigel

J Cell Physiol. 1995 Feb;162(2):213-24. doi: 10.1002/jcp.1041620207.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Capillaries / growth & development*
  • Capillaries / metabolism
  • Collagen*
  • Collagenases / biosynthesis
  • Drug Combinations
  • Endothelium, Vascular / cytology
  • Endothelium, Vascular / growth & development*
  • Endothelium, Vascular / metabolism
  • Epidermal Growth Factor / pharmacology
  • Gene Expression Regulation, Developmental / drug effects
  • Glycoproteins / biosynthesis
  • Humans
  • Kinetics
  • Laminin*
  • Matrix Metalloproteinase 9
  • Matrix Metalloproteinase Inhibitors
  • Metalloendopeptidases / antagonists & inhibitors
  • Peritoneum / cytology
  • Plasminogen Activator Inhibitor 1 / biosynthesis*
  • Protein Biosynthesis
  • Proteoglycans*
  • RNA, Messenger / biosynthesis
  • Tissue Inhibitor of Metalloproteinase-2
  • Tissue Inhibitor of Metalloproteinases
  • Tissue Plasminogen Activator / biosynthesis*
  • Tissue Plasminogen Activator / pharmacology
  • Transforming Growth Factor beta / physiology


  • Drug Combinations
  • Glycoproteins
  • Laminin
  • Matrix Metalloproteinase Inhibitors
  • Plasminogen Activator Inhibitor 1
  • Proteoglycans
  • RNA, Messenger
  • Tissue Inhibitor of Metalloproteinases
  • Transforming Growth Factor beta
  • matrigel
  • Tissue Inhibitor of Metalloproteinase-2
  • Epidermal Growth Factor
  • Collagen
  • Tissue Plasminogen Activator
  • Collagenases
  • Metalloendopeptidases
  • Matrix Metalloproteinase 9