Document Detail

Inhibition of Rho-Kinase Abrogates Migration of Human Transitional Cell Carcinoma Cells: Results of an in vitro Study.
MedLine Citation:
PMID:  21051874     Owner:  NLM     Status:  In-Data-Review    
Introduction: Migration of cells involves a complex signaling network. The aim of the present study was to elucidate the impact of Rho-kinase (ROK) on G protein-coupled receptor-induced migration of human transitional cell carcinoma cells in an in vitro experimental setting. Materials and Methods: Intracellular calcium concentration ([Ca(2+)](i)) was measured with the indicator dye Fura-2 in response to lysophosphatidic acid, thrombin and sphingosine-1-phosphate. Phospholipase C activity was determined in myo-[(3)H]inositol- (0.5 μCi/ml) labeled cells. Migration was performed using a Boyden chamber. Transient transfection of a dominant-negative mutant of ROK was done with calcium phosphate. For staining of actin filaments, tetramethylrhodamine isothiocyanate-conjugated phalloidin was used. Results: Lysophosphatidic acid, thrombin and sphingosine-1-phosphate cause increases in [Ca(2+)](i), cellular responses being accompanied by an enhancement of phospholipase C activity and sensitive to the G(i) inhibitor pertussis toxin. Agonists potently stimulated migration of T24 and J82 cells. Inhibition of Rho proteins by Clostridium difficile toxin B abrogated cell migration. Inhibition of ROK using HA1077 and Y-27632 mimicked the properties of toxin B. Expression of a ROK mutant drastically reduced migration. Conclusions: G protein-coupled receptors potently stimulated cell migration in T24 and J82 cells. Rho proteins and ROK play a pivotal role in this signaling cascade. Rho and ROK may be putative targets for new therapy options in bladder cancer.
Frank Vom Dorp; Harald Sanders; Christof Boergermann; Gerd Lümmen; Herbert Rübben; Karl H Jakobs; Martina Schmidt
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Publication Detail:
Type:  Journal Article     Date:  2010-11-04
Journal Detail:
Title:  Urologia internationalis     Volume:  86     ISSN:  1423-0399     ISO Abbreviation:  Urol. Int.     Publication Date:  2011  
Date Detail:
Created Date:  2011-03-22     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0417373     Medline TA:  Urol Int     Country:  Switzerland    
Other Details:
Languages:  eng     Pagination:  220-7     Citation Subset:  IM    
Copyright Information:
Copyright © 2010 S. Karger AG, Basel.
Institute for Pharmacology, University of Essen Medical School, Essen, Germany.
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