Inhibition of Rho-Kinase Abrogates Migration of Human Transitional Cell Carcinoma Cells: Results of an in vitro Study
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: I...
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| Veröffentlicht in: | Urologia internationalis 2011-01, Vol.86 (2), p.220-227 |
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| description | 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. |
| doi_str_mv | 10.1159/000321271 |
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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.</description><identifier>ISSN: 0042-1138</identifier><identifier>EISSN: 1423-0399</identifier><identifier>DOI: 10.1159/000321271</identifier><identifier>PMID: 21051874</identifier><language>eng</language><publisher>Basel, Switzerland</publisher><subject>1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives ; 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology ; Actins - chemistry ; Amides - pharmacology ; Bacterial Proteins - chemistry ; Bacterial Toxins - chemistry ; Calcium - chemistry ; Calcium - metabolism ; Carcinoma, Transitional Cell - enzymology ; Cell Line, Tumor ; Cell Movement ; Fura-2 - pharmacology ; Gene Expression Regulation, Neoplastic ; Humans ; In Vitro Techniques ; Lysophospholipids - chemistry ; Lysophospholipids - pharmacology ; Mutation ; Original Paper ; Pertussis Toxin - pharmacology ; Phalloidine - chemistry ; Protein Kinase Inhibitors - pharmacology ; Pyridines - pharmacology ; rho GTP-Binding Proteins - metabolism ; rho-Associated Kinases - antagonists & inhibitors ; rho-Associated Kinases - metabolism ; Type C Phospholipases - metabolism</subject><ispartof>Urologia internationalis, 2011-01, Vol.86 (2), p.220-227</ispartof><rights>2010 S. 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Karger AG, Basel.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c371t-b49da84c87a2d416648254a1d8bbbb43e1d7229b6265e8c52583a40ed10d8e143</citedby><cites>FETCH-LOGICAL-c371t-b49da84c87a2d416648254a1d8bbbb43e1d7229b6265e8c52583a40ed10d8e143</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,2429,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/21051874$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>vom Dorp, Frank</creatorcontrib><creatorcontrib>Sanders, Harald</creatorcontrib><creatorcontrib>Boergermann, Christof</creatorcontrib><creatorcontrib>Lümmen, Gerd</creatorcontrib><creatorcontrib>Rübben, Herbert</creatorcontrib><creatorcontrib>Jakobs, Karl H.</creatorcontrib><creatorcontrib>Schmidt, Martina</creatorcontrib><title>Inhibition of Rho-Kinase Abrogates Migration of Human Transitional Cell Carcinoma Cells: Results of an in vitro Study</title><title>Urologia internationalis</title><addtitle>Urol Int</addtitle><description>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.</description><subject>1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives</subject><subject>1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology</subject><subject>Actins - chemistry</subject><subject>Amides - pharmacology</subject><subject>Bacterial Proteins - chemistry</subject><subject>Bacterial Toxins - chemistry</subject><subject>Calcium - chemistry</subject><subject>Calcium - metabolism</subject><subject>Carcinoma, Transitional Cell - enzymology</subject><subject>Cell Line, Tumor</subject><subject>Cell Movement</subject><subject>Fura-2 - pharmacology</subject><subject>Gene Expression Regulation, Neoplastic</subject><subject>Humans</subject><subject>In Vitro Techniques</subject><subject>Lysophospholipids - chemistry</subject><subject>Lysophospholipids - pharmacology</subject><subject>Mutation</subject><subject>Original Paper</subject><subject>Pertussis Toxin - pharmacology</subject><subject>Phalloidine - chemistry</subject><subject>Protein Kinase Inhibitors - pharmacology</subject><subject>Pyridines - pharmacology</subject><subject>rho GTP-Binding Proteins - metabolism</subject><subject>rho-Associated Kinases - antagonists & inhibitors</subject><subject>rho-Associated Kinases - metabolism</subject><subject>Type C Phospholipases - metabolism</subject><issn>0042-1138</issn><issn>1423-0399</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo90M1LwzAYBvAgipvTg3eR3MRDNW-atqm3MdQNJ8Kc55I22RZdk5m0wv57s88cEgK_5-XlQegayANAkj8SQmIKNIMT1AVG44jEeX6KuoQwGgHEvIMuvP8mJOA8O0cdCiQBnrEuakdmoUvdaGuwneHJwkZv2givcL90di4a5fG7njtxEMO2FgZPnTB-mxJLPFDLcAlXaWNrsf36JzxRvl02fpMJAW3wn26cxZ9NK9eX6Gwmll5d7d8e-np5ng6G0fjjdTToj6MqzqCJSpZLwVnFM0ElgzRlnCZMgORlOCxWIDNK8zKlaaJ4ldCEx4IRJYFIroDFPXS3m7ty9rdVvilq7auwnzDKtr7gCac8VAJB3u9k5az3Ts2KldO1cOsCSLEpuTiWHOztfmpb1koe5aHVAG524Ee4uXJHsM__A0AJf3E</recordid><startdate>20110101</startdate><enddate>20110101</enddate><creator>vom Dorp, Frank</creator><creator>Sanders, Harald</creator><creator>Boergermann, Christof</creator><creator>Lümmen, Gerd</creator><creator>Rübben, Herbert</creator><creator>Jakobs, Karl H.</creator><creator>Schmidt, Martina</creator><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7X8</scope></search><sort><creationdate>20110101</creationdate><title>Inhibition of Rho-Kinase Abrogates Migration of Human Transitional Cell Carcinoma Cells: Results of an in vitro Study</title><author>vom Dorp, Frank ; Sanders, Harald ; Boergermann, Christof ; Lümmen, Gerd ; Rübben, Herbert ; Jakobs, Karl H. ; Schmidt, Martina</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c371t-b49da84c87a2d416648254a1d8bbbb43e1d7229b6265e8c52583a40ed10d8e143</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives</topic><topic>1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology</topic><topic>Actins - chemistry</topic><topic>Amides - pharmacology</topic><topic>Bacterial Proteins - chemistry</topic><topic>Bacterial Toxins - chemistry</topic><topic>Calcium - chemistry</topic><topic>Calcium - metabolism</topic><topic>Carcinoma, Transitional Cell - enzymology</topic><topic>Cell Line, Tumor</topic><topic>Cell Movement</topic><topic>Fura-2 - pharmacology</topic><topic>Gene Expression Regulation, Neoplastic</topic><topic>Humans</topic><topic>In Vitro Techniques</topic><topic>Lysophospholipids - chemistry</topic><topic>Lysophospholipids - pharmacology</topic><topic>Mutation</topic><topic>Original Paper</topic><topic>Pertussis Toxin - pharmacology</topic><topic>Phalloidine - chemistry</topic><topic>Protein Kinase Inhibitors - pharmacology</topic><topic>Pyridines - pharmacology</topic><topic>rho GTP-Binding Proteins - metabolism</topic><topic>rho-Associated Kinases - antagonists & inhibitors</topic><topic>rho-Associated Kinases - metabolism</topic><topic>Type C Phospholipases - metabolism</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>vom Dorp, Frank</creatorcontrib><creatorcontrib>Sanders, Harald</creatorcontrib><creatorcontrib>Boergermann, Christof</creatorcontrib><creatorcontrib>Lümmen, Gerd</creatorcontrib><creatorcontrib>Rübben, Herbert</creatorcontrib><creatorcontrib>Jakobs, Karl H.</creatorcontrib><creatorcontrib>Schmidt, Martina</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>MEDLINE - Academic</collection><jtitle>Urologia internationalis</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>vom Dorp, Frank</au><au>Sanders, Harald</au><au>Boergermann, Christof</au><au>Lümmen, Gerd</au><au>Rübben, Herbert</au><au>Jakobs, Karl H.</au><au>Schmidt, Martina</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Inhibition of Rho-Kinase Abrogates Migration of Human Transitional Cell Carcinoma Cells: Results of an in vitro Study</atitle><jtitle>Urologia internationalis</jtitle><addtitle>Urol Int</addtitle><date>2011-01-01</date><risdate>2011</risdate><volume>86</volume><issue>2</issue><spage>220</spage><epage>227</epage><pages>220-227</pages><issn>0042-1138</issn><eissn>1423-0399</eissn><abstract>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.</abstract><cop>Basel, Switzerland</cop><pmid>21051874</pmid><doi>10.1159/000321271</doi><tpages>8</tpages></addata></record> |
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| subjects | 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - analogs & derivatives 1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine - pharmacology Actins - chemistry Amides - pharmacology Bacterial Proteins - chemistry Bacterial Toxins - chemistry Calcium - chemistry Calcium - metabolism Carcinoma, Transitional Cell - enzymology Cell Line, Tumor Cell Movement Fura-2 - pharmacology Gene Expression Regulation, Neoplastic Humans In Vitro Techniques Lysophospholipids - chemistry Lysophospholipids - pharmacology Mutation Original Paper Pertussis Toxin - pharmacology Phalloidine - chemistry Protein Kinase Inhibitors - pharmacology Pyridines - pharmacology rho GTP-Binding Proteins - metabolism rho-Associated Kinases - antagonists & inhibitors rho-Associated Kinases - metabolism Type C Phospholipases - metabolism |
| title | Inhibition of Rho-Kinase Abrogates Migration of Human Transitional Cell Carcinoma Cells: Results of an in vitro Study |
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