Detachment of murine Balb/c 3T3 cells (or their oncogenic virus transformants or partially adhesion-defective variants) from the tissue culture substratum (coated with plasma fibronectin [pFn] with EGTA treatment leaves tight-focal adhesion sites and some close-contact adhesion sites on the undersurface of the cell as substratum-attached material (SAM). SAM is a considerable enrichment of the adhesive material of these cells and affords us the opportunity to examine the biochemical requirements for forming tight focal contacts and close contacts. Since SAM contains principally heparan sulfate proteoglycan (HS-PG), hyaluronate, and cell surface fibronectin (cFn), along with some chondroitin sulfate proteoglycan and cytoskeletal elements, we investigated and established evidence for binding of cFn or pFn to HS chains and the binding preferentially of CFn to HA. We will continue to study the properties of the two forms of Fn binding to these glycosaminoglycan-containing elements and the possible importance of these bindings in forming or altering the two types of adhesive contacts described above. Specifically, the binding of the Fn's to HS-PG, rather than free chains, will be examine in more detail. Second, the requirements and properties of binding of cFn to HA will be studied with the eventual goal of better understanding the simultaneous binding of HA and HS-PG to cFn and possible consequences. Third, we will attempt to isolate cFn:HS-Pg and/or cFn:HA complexes from the SAM's of these cells for characterization under various growth and attachment conditions. Finally, we will study the significance of Fn:GAG binding in substratum adhesion of these cells by comparing the adhesive responses of untransformed, transformed, or adhesion-variant cells attaching to substrata coated with pFn (or its specific binding domains) or with specific GAG-binding proteins. (A)

Agency
National Institute of Health (NIH)
Institute
National Cancer Institute (NCI)
Type
Research Project (R01)
Project #
5R01CA027755-05
Application #
3167803
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1981-01-01
Project End
1988-12-31
Budget Start
1985-01-01
Budget End
1985-12-31
Support Year
5
Fiscal Year
1985
Total Cost
Indirect Cost
Name
Case Western Reserve University
Department
Type
Schools of Medicine
DUNS #
077758407
City
Cleveland
State
OH
Country
United States
Zip Code
44106
Culp, L A; Lin, W C; Kleinman, N R (1999) Tagged tumor cells reveal regulatory steps during earliest stages of tumor progression and micrometastasis. Histol Histopathol 14:879-86
Kogerman, P; Sy, M S; Culp, L A (1998) Over-expression of human CD44s in murine 3T3 cells: selection against during primary tumorigenesis and selection for during micrometastasis. Clin Exp Metastasis 16:83-93
Culp, L A; Lin, W C; Kleinman, N R et al. (1998) Tumor progression, micrometastasis, and genetic instability tracked with histochemical marker genes. Prog Histochem Cytochem 33:XI-XV, 329-48
Culp, L A; Lin, W; Kleinman, N R et al. (1998) Earliest steps in primary tumor formation and micrometastasis resolved with histochemical markers of gene-tagged tumor cells. J Histochem Cytochem 46:557-68
Culp, L A; Sukenik, C N (1998) Cell type-specific modulation of fibronectin adhesion functions on chemically-derivatized self-assembled monolayers. J Biomater Sci Polym Ed 9:1161-76
Kogerman, P; Sy, M S; Culp, L A (1997) Overexpressed human CD44s promotes lung colonization during micrometastasis of murine fibrosarcoma cells: facilitated retention in the lung vasculature. Proc Natl Acad Sci U S A 94:13233-8
Kogerman, P; Sy, M S; Culp, L A (1997) Counter-selection for over-expressed human CD44s in primary tumors versus lung metastases in a mouse fibrosarcoma model. Oncogene 15:1407-16
Kogerman, P; Sy, M S; Culp, L A (1996) Oncogene-dependent expression of CD44 in Balb/c 3T3 derivatives: correlation with metastatic competence. Clin Exp Metastasis 14:73-82
Kogerman, P; Sy, M S; Culp, L A (1996) CD44 protein levels and its biological activity are regulated in Balb/c 3T3 fibroblasts by serum factors and by transformation with the ras but not with the sis oncogene. J Cell Physiol 169:341-9
Xia, P; Culp, L A (1995) Adhesion activity in fibronectin's alternatively spliced domain EDa (EIIIA): complementarity to plasma fibronectin functions. Exp Cell Res 217:517-27

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