Invasion of ras-Transformed Breast Epithelial Cells Depends on the Proteolytic Activity of Cysteine and Aspartic Proteinases

• DOI: 10.1515/bc.2001.104
• Publication Date: 2004-10-11T16:14:55Z
• Authors: A. Premzl; V. Puizdar; V. Zavanik-Bergant; N. Kopitar-Jerala; T.T. Lah; N. Katunuma; B.F. Sloane; V. Turk; J. Kos
• Keywords: Pepstatin; Cystatin; Cysteine Proteinase Inhibitors; Cathepsin L; Cathepsin H; Cathepsin L1; Proteolytic enzymes

It has been suggested that the lysosomal proteinases cathepsin B, L and D participate in tumour invasion metastasis. Whereas for cathepsins B role of active enzyme processes confirmed, was to support progression via its pro-peptide, rather than by proteolytic activity. In this study we have compared presence ras-transformed human breast epithelial cells (MCF-10A neoT) with their ability invade matrigel. cell line high expression all three detected immunofluorescence microscopy. The effect activity on studied adding various natural synthetic cysteine aspartic proteinase inhibitors. most effective compound chicken cystatin, a general inhibitor proteinases, (82.8+/-1.6% inhibition invasion), followed trans-epoxysuccinyl-L-leucylamido-(4-guanidino) butane (E-64). CLIK-148, specific L, showed lower cystatin E-64. Pepstatin A weakly inhibited invasion, whereas same molar concentrations squash (SQAPI)-like inhibitor, isolated from Cucurbita pepo, significant (65.7+/-1.8%). We conclude both activities are needed MCF-10A neoT vitro.