Rho-kinase inhibitor upregulates migration by altering focal adhesion formation via the Akt pathway in colon cancer cells.
Although Rho-kinase is reportedly implicated in carcinogenesis and the progression of human cancers, its precise mechanism has not been fully elucidated. We recently reported that Rho-kinase negatively regulates epidermal growth factor (EGF)-stimulated cancer progression in SW480 colon cancer cells. In the present study, we investigated the effect of Rho-kinase on the migration of SW480 colon cancer cells and the mechanism underlying the involvement of Rho-kinase. Interestingly, (R)-(+)-trans-N-(4-pyridyl)-4-(1-aminoethyl)-cyclohexanecarboxamide, 2HCl (Y27632), a specific inhibitor of Rho-kinase, dose-dependently enhanced cell migration. SW480 cells spontaneously release vascular endothelial growth factor (VEGF), however, Y27632 had little effect on its release. While Rho-kinase, which is generally phosphorylated in unstimulated cells, was clearly suppressed by Y27632, exogenous VEGF did not affect its phosphorylation. Immunofluorescence microscopy revealed that Y27632 caused a dramatic change in the localization of focal adhesion components, vinculin, phosphorylated caveolin-1 and tyrosine-phosphorylated proteins in SW480 cells. Furthermore, Akt inhibitor restored the loss of vinculin-stained focal adhesion formation induced by Y27632. We also observed similar effects for Y27632 on the migration and localization of focal adhesion components such as vinculin in another colon cancer cell line, HT29. Taken together, these results strongly suggest that Rho-kinase negatively regulates the migration of colon cancer cells by altering focal adhesion formation via the Akt pathway.
Adachi
Seiji
S
Department of Gastroenterology, Gifu University Graduate School of Medicine, Gifu 501-1194, Japan. seijiadachi0123@gmail.com
Yasuda
Ichiro
I
Nakashima
Masanori
M
Yamauchi
Takahiro
T
Yoshioka
Takashi
T
Okano
Yukio
Y
Moriwaki
Hisataka
H
Kozawa
Osamu
O
D000577:Amides / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects*; D003110:Colonic Neoplasms / Q000473:pathology*; D022001:Focal Adhesions / Q000187:drug effects* / Q000378:metabolism; D019073:HT29 Cells; D006801:Humans; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D021381:Protein Transport / Q000187:drug effects; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D011725:Pyridines / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects*; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism / Q000494:pharmacology; D054460:rho-Associated Kinases / Q000037:antagonists & inhibitors*
D000577:Amides; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D042461:Vascular Endothelial Growth Factor A; C108830:Y 27632; D051057:Proto-Oncogene Proteins c-akt; D054460:rho-Associated Kinases