Disruption of focal adhesion kinase slows transendothelial migration of AU-565 breast cancer cells.
Transendothelial migration of cancer cells from the vasculature into tissue stroma is a final step in the metastatic cascade, prior to formation of secondary tumors. Due to its role in 2-dimensional migration of cells on extracellular matrix proteins, we hypothesized that focal adhesion kinase (FAK) promotes transendothelial migration of cancer cells. AU-565 cells are weakly invasive metastatic breast adenocarcinoma cells that migrate through bovine lung microvessel endothelial cell monolayers. Electric cell-substrate impedance sensing detects a significant decrease in monolayer resistance upon addition of AU-565 cells. Immunofluorescence microscopy and filter-based migration assays demonstrate that this drop in resistance correlates with transendothelial migration. Transfection of AU-565 cells with FAK siRNA results in significantly diminished transendothelial migration of AU-565 cells within 15h. Expression of the dominant negative FAK inhibitor FAK-related non-kinase (FRNK) also results in delayed AU-565 transendothelial migration, whereas over-expression of wildtype FAK does not impact transendothelial migration substantially. These results demonstrate that FAK affects the rate of a key step in the metastatic cascade.
Earley
Sarah
S
Department of Biology, Center for Biotechnology and Interdisciplinary Studies, BCHM-2, Rensselaer Polytechnic Institute, 110 8th Street, Troy, NY 12180, USA.
Plopper
George E
GE
D000818:Animals; D001943:Breast Neoplasms / Q000201:enzymology* / Q000503:physiopathology / Q000648:ultrastructure; D002417:Cattle; D045744:Cell Line, Tumor; D002465:Cell Movement; D042783:Endothelial Cells / Q000648:ultrastructure; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000235:genetics; D006801:Humans; D011505:Protein-Tyrosine Kinases / Q000378:metabolism; D034622:RNA Interference
C113183:FAK-related nonkinase; D011505:Protein-Tyrosine Kinases; D051416:Focal Adhesion Protein-Tyrosine Kinases