Transendothelial Migration of Colon Carcinoma Cells Requires Expression of E-selectin by Endothelial Cells and Activation of Stress-activated Protein Kinase-2 (SAPK2/p38) in the Tumor Cells

• Published in: The Journal of biological chemistry 2001-09, Vol.276 (36), p.33762-33772
• Main Authors: Laferrière, Julie, Houle, François, Taher, Mohiuddin M., Valerie, Kristoffer, Huot, Jacques
• Format: Article
• Language: English
• Subjects: Cell Adhesion; Cell Movement; Cells, Cultured; Colonic Neoplasms - metabolism; Dose-Response Relationship, Drug; E-selectin; E-Selectin - biosynthesis; Endothelium - metabolism; Endothelium, Vascular - cytology; Enzyme Activation; Enzyme Inhibitors - pharmacology; Heat-Shock Proteins; HSP27 Heat-Shock Proteins; Humans; Imidazoles - pharmacology; Microscopy, Confocal; Mitogen-Activated Protein Kinase 1 - metabolism; Mitogen-Activated Protein Kinases - metabolism; Molecular Chaperones; Neoplasm Proteins - metabolism; Oxygen - metabolism; p38 Mitogen-Activated Protein Kinases; Phosphorylation; Plant Proteins; Precipitin Tests; Protein Binding; Pyridines - pharmacology; SAPK2 protein; stress-activated protein kinase-2; Time Factors; Transfection; Tumor Cells, Cultured; Tumor Necrosis Factor-alpha - metabolism; Umbilical Veins - metabolism
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.M008564200

Adhesion and migration of tumor cells on and through the vascular endothelium are critical steps of the metastatic invasion. We investigated the roles of E-selectin and of stress-activated protein kinase-2 (SAPK2/p38) in modulating endothelial adhesion and transendothelial migration of HT-29 colon carcinoma cells. Tumor necrosis factor α (TNFα) strongly increased the expression of E-selectin in human umbilical vein endothelial cells (HUVEC). This effect was independent of the activation of SAPK2/p38 induced by TNFα. Adhesion of HT-29 cells on a monolayer of HUVEC pretreated with TNFα was dependent on E-selectin expression but was independent of SAPK2/p38 activity of both HUVEC and tumor cells. The adhesion of HT-29 cells to E-selectin-expressing HUVEC led to the activation of SAPK2/p38 in the tumor cells as reflected by the increased phosphorylation of the actin-polymerizing factor HSP27 by mitogen-activated protein kinase 2/3, a direct target of SAPK2/p38. Moreover, a recombinant E-selectin/Fc chimera quickly increased the activation of SAPK2/p38 in HT-29 cells. Blocking the increased activity of SAPK2/p38 of HT-29 cells by SB203580 or by expressing a dominant negative form of SAPK2/p38 inhibited their transendothelial migration. Similarly, HeLa cells stably expressing a kinase-inactive mutant of SAPK2/p38 showed a decreased capacity to cross a layer of HUVEC. Overall, our results suggest that the regulation of transendothelial migration of tumor cells involves two essential steps as follows: adhesion to the endothelium through adhesion molecules, such as E-selectin, and increased motogenic potential through adhesion-mediated activation of the SAPK2/p38 pathway.