The metastatic potential of tumor cells is the most important determinant for the prognosis of neoplastic disease. Among the molecular steps required for successful metastasis are several discrete events that involve the invasion of tumor cells through connective tissue barriers. Cell invasiveness concerns adhesion to and proteolysis of extracellular matrices and basement membranes, as well as cell motility. This proposal focuses on plasma membrane-associated proteases and adhesion molecules of human malignant melanoma cells that are important in mediating the invasiveness and metastatic potential of these cells. Recent work from this laboratory identifies several novel membrane proteases from human malignant melanoma cells, chicken embryonic cells, chicken transformed cells, and chicken embryos. Unique cell surface structures called invadopodia are present on cells expressing the malignant phenotype and have been shown to be involved in the invasiveness of cells. The invadopodia are sites of active tyrosine-phosphorylation, localization of beta1 integrins, and active degradation of the extracellular matrix by membrane proteases. The goals of this proposal are (1) to determine the biochemical nature of membrane proteases by the combined approaches of protease activity assays, cellular localization by monoclonal antibody production, sub-cellular fractionation, inhibitor- and antibody-affinity chromatography, protein biochemistry including microsequencing, and molecular cloning, (2) to determine how membrane proteases interact with other proteases to activate degradation, and their role in the expression of cell invasiveness, (3) to investigate the control mechanisms of cell invasion, particularly focusing on the direct or indirect interaction of membrane proteases with putative motility factors including the laminin A chain peptide, autocrine motility factor, acidic fibroblast growth factor, and interleukin-I during the induction of invadopodia and with the human integrins that localize to invadopodia during the stabilization of invasion-associated membranes, and (4) to determine the expression and localization of protease and adhesion molecules in experimental invasion and metastasis models and human tumors by scanning confocal microscopy and immuno-electron microscopy.

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
2R01CA039077-09
Application #
3177844
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1984-07-01
Project End
1996-11-30
Budget Start
1992-01-10
Budget End
1992-11-30
Support Year
9
Fiscal Year
1992
Total Cost
Indirect Cost
Name
Georgetown University
Department
Type
Schools of Medicine
DUNS #
049515844
City
Washington
State
DC
Country
United States
Zip Code
20057
Tulley, Shaun; Chen, Wen-Tien (2014) Transcriptional regulation of seprase in invasive melanoma cells by transforming growth factor-? signaling. J Biol Chem 289:15280-96
Javidroozi, Mazyar; Zucker, Stanley; Chen, Wen-Tien (2012) Plasma seprase and DPP4 levels as markers of disease and prognosis in cancer. Dis Markers 32:309-20
Lu, Janice; Fan, Tina; Zhao, Qiang et al. (2010) Isolation of circulating epithelial and tumor progenitor cells with an invasive phenotype from breast cancer patients. Int J Cancer 126:669-83
Fan, Tina; Zhao, Qiang; Chen, John J et al. (2009) Clinical significance of circulating tumor cells detected by an invasion assay in peripheral blood of patients with ovarian cancer. Gynecol Oncol 112:185-91
Kennedy, Alanna; Dong, Huan; Chen, Donghai et al. (2009) Elevation of seprase expression and promotion of an invasive phenotype by collagenous matrices in ovarian tumor cells. Int J Cancer 124:27-35
Freudenberg, Jaclyn A; Chen, Wen-Tien (2007) Induction of Smad1 by MT1-MMP contributes to tumor growth. Int J Cancer 121:966-77
Ghersi, Giulio; Zhao, Qiang; Salamone, Monica et al. (2006) The protease complex consisting of dipeptidyl peptidase IV and seprase plays a role in the migration and invasion of human endothelial cells in collagenous matrices. Cancer Res 66:4652-61
Chen, Donghai; Kennedy, Alanna; Wang, Jaw-Yuan et al. (2006) Activation of EDTA-resistant gelatinases in malignant human tumors. Cancer Res 66:9977-85
Chen, Wen-Tien; Kelly, Thomas (2003) Seprase complexes in cellular invasiveness. Cancer Metastasis Rev 22:259-69
Chen, Wen-Tien; Kelly, Thomas; Ghersi, Giulio (2003) DPPIV, seprase, and related serine peptidases in multiple cellular functions. Curr Top Dev Biol 54:207-32

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