Expression of tissue-type plasminogen activator and its inhibitor couples with development of capillary network by human microvascular endothelial cells on matrigel
| Autor: | Ken-ichi Ito, Yosaburo Shibata, Akira Sugenoya, Michihiko Kuwano, Mayumi Ono, Shin Ushiro, Akio Kuraoka, Masahiro Ryuto |
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| Rok vydání: | 1995 |
| Předmět: |
Physiology
Clinical Biochemistry Matrix Metalloproteinase Inhibitors Transforming Growth Factor beta Epidermal growth factor Plasminogen Activator Inhibitor 1 Gene expression Humans Collagenases RNA Messenger Northern blot Glycoproteins Tissue Inhibitor of Metalloproteinase-2 Messenger RNA Matrigel Epidermal Growth Factor biology Gene Expression Regulation Developmental Metalloendopeptidases Tissue Inhibitor of Metalloproteinases Cell Biology Tissue inhibitor of metalloproteinase Molecular biology Capillaries Drug Combinations Kinetics Matrix Metalloproteinase 9 Protein Biosynthesis Tissue Plasminogen Activator biology.protein Proteoglycans Collagen Endothelium Vascular Laminin Peritoneum Antibody Plasminogen activator |
| Zdroj: | Journal of Cellular Physiology. 162:213-224 |
| ISSN: | 1097-4652 0021-9541 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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