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Am J Physiol Cell Physiol 279: C1472-C1482, 2000;
0363-6143/00 $5.00
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Vol. 279, Issue 5, C1472-C1482, November 2000

Differential expression of cell-cell adhesion proteins and cyclin D in MEK1-transdifferentiated MDCK cells

Ingrid Marschitz1, Judith Lechner1, Irene Mosser1, Martina Dander1, Roberto Montesano2, and Herbert Schramek1

1 Department of Physiology, University of Innsbruck, A-6010 Innsbruck, Austria; and 2 Department of Morphology, University Medical Center, CH-1211 Geneva 4, Switzerland

Overexpression of a constitutively active mutant of the mitogen-activated protein kinase kinase MEK1 (caMEK1) in epithelial Madin-Darby canine kidney (MDCK)-C7 cells disrupts morphogenesis, induces an invasive phenotype, and is associated with a reduced rate of cell proliferation. The role of cell-cell adhesion molecules and cell cycle proteins in these processes, however, has not been investigated. We now report loss of E-cadherin expression as well as a marked reduction of beta - and alpha -catenin expression in transdifferentiated MDCK-C7 cells stably expressing caMEK1 (C7caMEK1) compared with epithelial mock-transfected MDCK-C7 (C7Mock1) cells. At least part of the remaining alpha -catenin was coimmunoprecipitated with beta -catenin, whereas no E-cadherin was detected in beta -catenin immunoprecipitates. In both cell types, the proteasome-specific protease inhibitors N-acetyl-Leu-Leu-norleucinal (ALLN) and lactacystin led to a time-dependent accumulation of beta -catenin, including the appearance of high-molecular-weight beta -catenin species. Quiescent as well as serum-stimulated C7caMEK1 cells showed a higher cyclin D expression than epithelial C7Mock1 cells. The MEK inhibitor U-0126 inhibited extracellular signal-regulated kinase phosphorylation and cyclin D expression in C7caMEK1 cells and almost abolished their already reduced cell proliferation rate. We conclude that the transdifferentiated and invasive phenotype of C7caMEK1 cells is associated with a diminished expression of proteins involved in cell-cell adhesion. Although beta -catenin expression is reduced, C7caMEK1 cells show a higher expression of U-0126-sensitive cyclin D protein.

extracellular signal-regulated kinase; differentiation; proliferation; E-cadherin; beta -catenin


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