The electrical resistance breakdown assay determines the role of proteinases in tumor cell invasion

The electrical resistance breakdown of the Madin-Darby canine kidney (MDCK) cell monolayer provides a continuous assay system for cancer invasion that detects functional changes before morphological alterations. In this study, we address the question of whether physical contact between tumor cell an...

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Bibliographic Details
Published in:American journal of physiology. Renal physiology 2002-08, Vol.283 (2), p.F319-F327
Main Authors: Ludwig, Thomas, Ossig, Rainer, Graessel, Susanne, Wilhelmi, Marianne, Oberleithner, Hans, Schneider, Stefan W
Format: Article
Language:English
Subjects:
Animals
Basement Membrane - enzymology
Cell-Free System
Coculture Techniques
Collagenases - metabolism
Electric Impedance
Exocytosis - physiology
Humans
Hydroxamic Acids
Indoles - pharmacology
Kidney - cytology
Matrix Metalloproteinase Inhibitors
Matrix Metalloproteinases - metabolism
Melanoma
Neoplasm Invasiveness
Protease Inhibitors - pharmacology
Tumor Cells, Cultured - enzymology
Tumor Cells, Cultured - pathology
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Summary:The electrical resistance breakdown of the Madin-Darby canine kidney (MDCK) cell monolayer provides a continuous assay system for cancer invasion that detects functional changes before morphological alterations. In this study, we address the question of whether physical contact between tumor cell and epithelial monolayer is a prerequisite for tumor cell invasion. When human melanoma cells were seeded directly (i.e., physical contact) on top of an electrically tight epithelial cell layer (5,800 +/- 106 Omega x cm2), electrical monolayer leakage led to an 18 +/- 3% reduction of transepithelial electrical resistance within 24 h. However, when melanoma cells were seeded close to the basolateral surface of the epithelial cell monolayer but separated by a filter membrane (i.e., no physical contact), electrical leakage occurred even more quickly (42 +/- 3% reduction in 24 h). Atomic force microscopy detected discrete structural changes between cells. Electrical leakage was effectively blocked by alpha2-macroglobulin or ilomastat, inhibitors of matrix metalloproteinases. We conclude that exocytosis of soluble proteases causes electrical breakdown of the MDCK monolayer, independently of physical contact between tumor cells and the monolayer.
ISSN:1931-857X
1522-1466
DOI:10.1152/ajprenal.00327.2001