This research focuses on routine use of the extracellular matrix (ECM) produced by cultured bovine endothelial cells as a naturally produced substrate adequate for sustaining adhesion, growth, and differentiation of human epithelial cells. Using this substrate and serum-free medium supplemented with HDL, we have developed conditions for the initiation and maintenance in monolayer culture of normal and carcinomatous human breast, bladder, and ovarian epithelial cells, as well as of normal human upper respiratory tract epithelium and first trimester decidual cells. The ECM has been used for screening the efficiency of various cytotoxic drugs in preventing the reimplantation of tumor cells to exposed areas of the bladder submucosa following transurethral resection of the primary tumor. Various chemical modifications of the ECM specifically affected its induction of cell attachment, proliferation, and differentiation, and various cell types responded differently to a given modification of the ECM. Different constituents of the ECM may therefore be held responsible for its induction of specific biological activities. The ECM produced by corneal endothelial cells and chicken embryo fibroblasts was compared. Vascular endothelial cells were stimulated to proliferate by both matrices but when cultured on the fibroblastic matrix they grew on top of each other and failed to adopt the monolayer configuration characteristic of endothelial cells cultured on a basement membrane-like ECM. A highly metastatic subline (ESb) of mouse lymphoma cells exhibited a much higher ability to degrade heparan sulfate than its non-metastatic parental subline (Eb). Degradation of ECM-bound, but not soluble, heparan sulfate proteoglycan proceeded in a sequential manner and was greatly facilitated in the presence of a serine protease which produced a more accessible substrate for further cleavage by the heparanase. Such a role was fulfilled by the ESb plasminogen activator via the generation of plasmin from plasminogen. A similar cascade mechanism was observed with other cell types whether normal or malignant. Heparanase activity was elaborated by normal platelets and granulocytes, and upon specific or nonspecific activation of normal macrophages and T lymphocytes. Antibasic protein (BP) lymphocytes responded to BP presented by ECM by a markedly enhanced heparanase activity, in correlation with their ability to extravasate the blood brain barrier. (A)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
3R01CA030289-06S1
Application #
3169201
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1981-09-30
Project End
1988-06-30
Budget Start
1986-12-01
Budget End
1988-06-30
Support Year
6
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Hadassah-Hebrew University Medical Center
Department
Type
DUNS #
600063937
City
Jerusalem
State
Country
Israel
Zip Code
91120
Benezra, M; Vlodavsky, I; Bar-Shavit, R (1993) Prothrombin conversion to thrombin by plasminogen activator residing in the subendothelial extracellular matrix. Semin Thromb Hemost 19:405-11
Korner, G; Bjornsson, T D; Vlodavsky, I (1993) Extracellular matrix produced by cultured corneal and aortic endothelial cells contains active tissue-type and urokinase-type plasminogen activators. J Cell Physiol 154:456-65
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Ishai-Michaeli, R; Svahn, C M; Weber, M et al. (1992) Importance of size and sulfation of heparin in release of basic fibroblast growth factor from the vascular endothelium and extracellular matrix. Biochemistry 31:2080-8
Vlodavsky, I; Ishai-Michaeli, R; Mohsen, M et al. (1992) Modulation of neovascularization and metastasis by species of heparin. Adv Exp Med Biol 313:317-27
Bashkin, P; Neufeld, G; Gitay-Goren, H et al. (1992) Release of cell surface-associated basic fibroblast growth factor by glycosylphosphatidylinositol-specific phospholipase C. J Cell Physiol 151:126-37
Benezra, M; Vlodavsky, I; Yayon, A et al. (1992) Reversal of basic fibroblast growth factor-mediated autocrine cell transformation by aromatic anionic compounds. Cancer Res 52:5656-62
Benezra, M; Vlodavsky, I; Bar-Shavit, R (1992) Thrombin enhances degradation of heparan sulfate in the extracellular matrix by tumor cell heparanase. Exp Cell Res 201:208-15
Vlodavsky, I; Eldor, A; Haimovitz-Friedman, A et al. (1992) Expression of heparanase by platelets and circulating cells of the immune system: possible involvement in diapedesis and extravasation. Invasion Metastasis 12:112-27

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