Suppression of matrix metalloproteinase activity by SI-27: detection by a new activity assay with S-2444, a specific chromogenic peptide

• Published in: Journal of neuro-oncology Vol. 58; no. 1; pp. 1 - 11
• Main Authors: YOSHIDA, Daizo, WATANABE, Kunihiro, NOHA, Masahiro, TAKAHASHI, Hiroshi, TERAMOTO, Akira
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.05.2002; Springer Nature B.V
• Subjects: Antineoplastic agents; Biological and medical sciences; Chemotherapy; Chromogenic Compounds; Collagen - metabolism; Dose-Response Relationship, Drug; Glioma - enzymology; Humans; Isoenzymes - antagonists & inhibitors; Isoenzymes - genetics; Isoenzymes - metabolism; Matrix Metalloproteinase Inhibitors; Matrix Metalloproteinases - genetics; Matrix Metalloproteinases - metabolism; Medical sciences; Neurology; Oligopeptides - administration & dosage; Oligopeptides - pharmacology; Pharmacology. Drug treatments; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger - metabolism; Tissue Inhibitor of Metalloproteinase-1 - antagonists & inhibitors; Tissue Inhibitor of Metalloproteinase-1 - genetics; Tissue Inhibitor of Metalloproteinase-1 - metabolism; Tissue Inhibitor of Metalloproteinase-2 - antagonists & inhibitors; Tissue Inhibitor of Metalloproteinase-2 - genetics; Tissue Inhibitor of Metalloproteinase-2 - metabolism; Tumor Cells, Cultured; Tumors of the nervous system. Phacomatoses; Antineoplastic agent; Human; Cell proliferation; Cell culture; Nervous system diseases; Enzyme; In vitro; Metalloproteins; Dose activity relation; Angiogenesis; Peptidases; Glioma; Central nervous system disease; Hydrolases; Tumor; Extracellular matrix; Pharmacokinetics
• ISSN: 0167-594X; 1573-7373
• DOI: 10.1023/A:1015894513482

We have previously reported on the anti-invasive and angiosuppressive effects of SI-27, an anti-matrix metalloproteinase (MMP) agent. The molecular mechanism of its anti-MMP action, however, has not yet been determined. The purpose of this study was to investigate the effects of SI-27 on MMP- 1, -2, -3, -9, and TIMP-1, -2 secreted by human glioma cell lines (U87MG, U251MG, U373MG, and Y98G). When cells were exposed to non-cytotoxic concentrations of SI-27 (preliminarily determined by the MTT assay), expressions of mRNAs for the enzymes was not inhibited. For an MMP activity assay, we employed the fact that active MMPs could cleave modified pro-urokinase to form active urokinase, which then acted on S-2444 peptide to create a chromogenic product. Secretion of all pro-MMPs from glioma cells was not significantly reduced by SI-27. However, activation of pro-MMPs was significantly inhibited in a dose-dependent manner ((IC50 values for MMP-2; U87MG, 3.5 microg/ml; U25 IMG, 4.2 microg/ml; U373MG, 4.8 microg/ml; Y98G, 4.0 degreesg/ml); (IC50 values for MMP-9; 251MG, 7.2 microg/ml, U373MG, 2.8 microg/ml). In addition, active MMPs were not inhibited by SI-27. These findings were supported by zymographic analysis and by collagenolysis assay data. TIMP-1 and -2 were also not inactivated by SI-27. These findings suggest that SI-27 targets the activation process of pro-MMP. S-2444, a specific chromogenic peptide, was useful for quantitative analysis of the activity of MMP subtypes in this study.