Abstract
Changes in the expression of several genes play critical roles in cell growth and tumor transformation. A number of proteases are increased in some tumors, and the level of these enzymes correlates with the metastatic potential of several cancer cell lines. Stromelysin, with the widest substrate specificity, can degrade the extracellular matrix conferring metastatic potential to tumor cells. The mechanisms whereby growth factors and oncogenes control the expression of stromelysin are beginning to be characterized. In the study shown here we also identify a region in the stromelysin promoter which is involved in the induction of stromelysin in response to platelet-derived growth factor, phosphatidylcholine-hydrolyzing phospholipase C, and ras oncogene. Our results are consistent with the notion that platelet-derived growth factor/phosphatidylcholine-hydrolyzing phospholipase C induces stromelysin gene expression through a phorbol myristate acetate/protein kinase C-independent mechanism by acting through elements in the stromelysin promoter distinct from the 12-O-tetradecanoylphorbol-13-acetate-responsive element.
Highlights
Changes in the expression of several genesplay crit- the level of these enzymes correlates with themetastatic ical roles in cell growth and tumor transformation
Number ofproteases are increased in some tumors, and Matrixdegradationrequirestheaction of these specific the level of these enzymes correlates with the metastatic potential of several cancer cell lines
In the study shown here we identify a region in the stromelysin promoter which is involved in the induction of stromelysin in response to platelet-derived growth factor,phosphatidylcholine-hydrolyzing phospholipase C, and ras oncogene
Summary
Cell Culturc>s-Human fibroblasts prepared from explantosf infant foreskin hy standardtechniqueswereculturedandmaintained as descrihed [21] in Dulbecco's modified Eagle's medium supplemented with 10'6 (v/v) fetal bovine serum, penicillin (100 units/ml), strepbranes (Amicon Corp., Lexington, MA). The enzyme was purified to complete homogeneity as confirmed by SDS-polyacrylamide gel electrophoresis followed by silver staining. The specific activity of the purified enzyme was1.5 units/pg [34].All chromatographic materials were from Pharmacia (Uppsala, Sweden). Tomycin (100 pg/ml), and mM bglutamine in standard tissue culture flasks in a humidified air/CO?
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