The importance of collagenases in tumor invasion and metastasis is well establised but mechanisms able to control the activity of these enzymes in tumors have not yet been explored. Studies of mechanisms of inhibition of these enzymes may be of importance to prevent tumor invasion. We have recently shown that bovine endothelial cells inhibit the collagenolytic activities normally displayed by human tumor cell lines. Subsequently we found 2 types of collagenase inhibitors in serum free medium conditioned by endothelial cells. Whereas one of these inhibitors with a molecular weight of 28,500 is similar to the ubiquitous tissue inhibitor of metalloproteinase (TIMP), the second has a molecular weight of approximately 70,000 and may be unique to endothelial cells. The role of these inhibitors in tumor invasion and metastasis will be investigated. The high molecular weight inhibitor will be isolated and purified from endothelial cell conditioned medium using various chromatographic procedures and its inhibitory activity against various collagenases including type IV specific collogenase will be tested. This will allow further comparision with other known inhibitors of collagenases. The low molecular weight inhibitor (TIMP) will be obtained as a recombinant protein. The ability of the purified high molecular weight and the recombinant TIMP to suppress the degradative and invasive activities of human tumor cells in vitro will then be explored. Rabbit polyclonal antibodies against these inhibitors will be raised and used to study their production by bovine endothelial cells of various origins (venous, arterial, capillary). The effect of phorbol diesters and cytochalasin D on the production of these inhibitors by endothelial cells will be investigated and the ability of tumor cells to secrete soluble factors influencing their biosynthesis by endothelial cells will be analyzed. These studies, will bring important and new information on the inhibition and modulation of tumor cell invasion and/or metastasis by endothelial cells.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
1R01CA042919-01A1
Application #
3184626
Study Section
Pathobiochemistry Study Section (PBC)
Project Start
1987-05-01
Project End
1990-04-30
Budget Start
1987-05-01
Budget End
1988-04-30
Support Year
1
Fiscal Year
1987
Total Cost
Indirect Cost
Name
Children's Hospital of Los Angeles
Department
Type
DUNS #
094878337
City
Los Angeles
State
CA
Country
United States
Zip Code
90027
Blavier, Laurence; Lazaryev, Alisa; Shi, Xiang-He et al. (2010) Stromelysin-1 (MMP-3) is a target and a regulator of Wnt1-induced epithelial-mesenchymal transition (EMT). Cancer Biol Ther 10:198-208
Wall, Steven J; Zhong, Zhi-Duan; DeClerck, Yves A (2007) The cyclin-dependent kinase inhibitors p15INK4B and p21CIP1 are critical regulators of fibrillar collagen-induced tumor cell cycle arrest. J Biol Chem 282:24471-6
Jodele, Sonata; Blavier, Laurence; Yoon, Janet M et al. (2006) Modifying the soil to affect the seed: role of stromal-derived matrix metalloproteinases in cancer progression. Cancer Metastasis Rev 25:35-43
Wall, Steven J; Werner, Erica; Werb, Zena et al. (2005) Discoidin domain receptor 2 mediates tumor cell cycle arrest induced by fibrillar collagen. J Biol Chem 280:40187-94
Wall, Steven J; Jiang, Yong; Muschel, Ruth J et al. (2003) Meeting report: Proteases, extracellular matrix, and cancer: an AACR Special Conference in Cancer Research. Cancer Res 63:4750-5
Blavier, L; Lazaryev, A; Groffen, J et al. (2001) TGF-beta3-induced palatogenesis requires matrix metalloproteinases. Mol Biol Cell 12:1457-66
Zhong, Z D; Hammani, K; Bae, W S et al. (2000) NF-Y and Sp1 cooperate for the transcriptional activation and cAMP response of human tissue inhibitor of metalloproteinases-2. J Biol Chem 275:18602-10
DeClerck, Y A (2000) Interactions between tumour cells and stromal cells and proteolytic modification of the extracellular matrix by metalloproteinases in cancer. Eur J Cancer 36:1258-68
Henriet, P; Zhong, Z D; Brooks, P C et al. (2000) Contact with fibrillar collagen inhibits melanoma cell proliferation by up-regulating p27KIP1. Proc Natl Acad Sci U S A 97:10026-31
Blavier, L; Henriet, P; Imren, S et al. (1999) Tissue inhibitors of matrix metalloproteinases in cancer. Ann N Y Acad Sci 878:108-19

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