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Am J Physiol Renal Physiol 283: F319-F327, 2002. First published February 26, 2002; doi:10.1152/ajprenal.00327.2001
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Vol. 283, Issue 2, F319-F327, August 2002

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

Thomas Ludwig1, Rainer Ossig1, Susanne Graessel2, Marianne Wilhelmi1, Hans Oberleithner1, and Stefan W. Schneider1

Institutes of 1 Physiology and 2 Biochemistry, University of Münster, D-48149 Münster, Germany

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  · 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 alpha 2-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.

basement membrane; exocytosis; melanoma cells; ilomastat; matrix metalloproteinases


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