Cysteine cathepsins are central contributors of invasion by cultured adenosylmethionine decarboxylase-transformed rodent fibroblasts.

Abstract

Adenosylmethionine decarboxylase (AdoMetDC), a key enzyme in the biosynthesis of polyamines, is often up-regulated in cancers. We have demonstrated previously that overexpression of AdoMetDC alone is sufficient to transform NIH 3T3 cells and induce highly invasive tumors in nude mice. Here, we studied the transformation-specific alterations in gene expression induced by AdoMetDC by using cDNA microarray and two-dimensional electrophoresis technologies. We specifically tried to identify the secreted proteins contributing to the high invasive activity of the AdoMetDC-transformed cells. We found a significant increase in the expression and secretion of procathepsin L, which was cleaved and activated in the presence of glycosaminoglycans (heparin), and a smaller increase in cathepsin B. Inhibition of the cathepsin L and B activity by specific peptide inhibitors abrogated the invasive capacity of the AdoMetDC transformants in Matrigel. The transformed cells also showed a small increase in the activity of gelatin-degrading matrix metalloproteinases (MMPs) and urokinase-type plasminogen activator activities, neither of which was sensitive to the inhibitors of cathepsin L and B. Furthermore, the invasive potency of the transformed cells remained unaffected by specific inhibitors of MMPs. The results suggest that cysteine cathepsins are the main proteases contributing to the high invasiveness of the AdoMetDC-transformed cells and that the invasion potential is largely independent of activation of the MMPs.

• Authors: Kirsi Ravanko, Kristiina Järvinen, Jari Helin, Nisse Kalkkinen, Erkki Hölttä
• Journal: Cancer research (Cancer Res) Vol. 64 Issue 24 Pg. 8831-8 (Dec 15 2004) ISSN: 0008-5472 [Print] United States
• PMID: 15604241 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• RNA, Messenger
• Cathepsins
• Urokinase-Type Plasminogen Activator
• Cysteine Endopeptidases
• Cathepsin B
• CTSL protein, human
• Cathepsin L
• Ctsl protein, mouse
• Ctsl protein, rat
• Matrix Metalloproteinases
• Adenosylmethionine Decarboxylase

Topics

• Adenosylmethionine Decarboxylase
• Animals
• Cathepsin B (metabolism)
• Cathepsin L
• Cathepsins (genetics, metabolism, physiology)
• Cell Movement (physiology)
• Cell Transformation, Neoplastic (genetics, metabolism)
• Cysteine Endopeptidases
• Endothelium, Vascular (cytology, enzymology)
• Enzyme Activation
• Fibroblasts (enzymology, pathology)
• Humans
• Immunoblotting
• Matrix Metalloproteinases (genetics, metabolism, physiology)
• Mice
• NIH 3T3 Cells
• Oligonucleotide Array Sequence Analysis
• RNA, Messenger (biosynthesis, genetics)
• Rats
• Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
• Transfection
• Urokinase-Type Plasminogen Activator (metabolism, physiology)