Role of Rho small GTPases in meniscus cells
ABSTRACT We previously reported that mechanical stretch regulates Sry‐type HMG box (SOX) 9‐dependent α1(II) collagen (COL2A1) expression in inner meniscus cells. This study examined the role of the small Rho guanosine 5' triphosphatase Rac1 and Rho‐associated kinase (ROCK) in the regulation of...
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| Veröffentlicht in: | Journal of orthopaedic research 2014-11, Vol.32 (11), p.1479-1486 |
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| container_title | Journal of orthopaedic research |
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| creator | Kanazawa, Tomoko Furumatsu, Takayuki Matsumoto-Ogawa, Emi Maehara, Ami Ozaki, Toshifumi |
| description | ABSTRACT
We previously reported that mechanical stretch regulates Sry‐type HMG box (SOX) 9‐dependent α1(II) collagen (COL2A1) expression in inner meniscus cells. This study examined the role of the small Rho guanosine 5' triphosphatase Rac1 and Rho‐associated kinase (ROCK) in the regulation of stretch‐induced SOX9 gene expression in cultured human inner meniscus cells. COL2A1 and SOX9 gene expression was assessed by real‐time PCR after application of uni‐axial cyclic tensile strain (CTS) in the presence or absence of ROCK and Rac1 inhibitors. The subcellular localization of SOX9 and the Rac1 effector cyclic AMP response element‐binding protein (CREB), the phosphorylation state of SOX9, Rac1 activation, and the binding of CREB to the SOX9 promoter were assessed. CTS increased the expression of COL2A1 and SOX9, which was suppressed by inhibition of Rac1. ROCK inhibition enhanced COL2A1 and SOX9 gene expression in the absence of CTS. CTS stimulated the nuclear translocation and phosphorylation of SOX9, and increased Rac1 activation. CTS also increased the binding of CREB to the SOX9 promoter. The results suggest that mechanical stretch‐dependent upregulation of SOX9 by CREB in inner meniscus cells depends on the antagonistic activities of ROCK and Rac1. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:1479–1486, 2014. |
| doi_str_mv | 10.1002/jor.22703 |
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We previously reported that mechanical stretch regulates Sry‐type HMG box (SOX) 9‐dependent α1(II) collagen (COL2A1) expression in inner meniscus cells. This study examined the role of the small Rho guanosine 5' triphosphatase Rac1 and Rho‐associated kinase (ROCK) in the regulation of stretch‐induced SOX9 gene expression in cultured human inner meniscus cells. COL2A1 and SOX9 gene expression was assessed by real‐time PCR after application of uni‐axial cyclic tensile strain (CTS) in the presence or absence of ROCK and Rac1 inhibitors. The subcellular localization of SOX9 and the Rac1 effector cyclic AMP response element‐binding protein (CREB), the phosphorylation state of SOX9, Rac1 activation, and the binding of CREB to the SOX9 promoter were assessed. CTS increased the expression of COL2A1 and SOX9, which was suppressed by inhibition of Rac1. ROCK inhibition enhanced COL2A1 and SOX9 gene expression in the absence of CTS. CTS stimulated the nuclear translocation and phosphorylation of SOX9, and increased Rac1 activation. CTS also increased the binding of CREB to the SOX9 promoter. The results suggest that mechanical stretch‐dependent upregulation of SOX9 by CREB in inner meniscus cells depends on the antagonistic activities of ROCK and Rac1. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:1479–1486, 2014.</description><identifier>ISSN: 0736-0266</identifier><identifier>EISSN: 1554-527X</identifier><identifier>DOI: 10.1002/jor.22703</identifier><identifier>PMID: 25130858</identifier><language>eng</language><publisher>United States: Blackwell Publishing Ltd</publisher><subject>Active Transport, Cell Nucleus ; Animals ; Arthroplasty, Replacement, Knee ; Biomechanical Phenomena ; Cartilage - enzymology ; Collagen Type II - metabolism ; CREB ; Cyclic AMP Response Element-Binding Protein - metabolism ; Gene Expression Regulation, Enzymologic ; Humans ; Knee - physiology ; Mechanical stretch ; Meniscus ; Phosphorylation ; Promoter Regions, Genetic ; Rac1 ; rac1 GTP-Binding Protein - metabolism ; Rats ; rho GTP-Binding Proteins - metabolism ; ROCK ; SOX9 ; SOX9 Transcription Factor - metabolism ; Stress, Mechanical ; Tensile Strength</subject><ispartof>Journal of orthopaedic research, 2014-11, Vol.32 (11), p.1479-1486</ispartof><rights>2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c4643-f9fb5f5219f921f2bc94554f57256f169f43722f0ad858e5e71f62467204a2613</citedby><cites>FETCH-LOGICAL-c4643-f9fb5f5219f921f2bc94554f57256f169f43722f0ad858e5e71f62467204a2613</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1002%2Fjor.22703$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1002%2Fjor.22703$$EHTML$$P50$$Gwiley$$H</linktohtml><link.rule.ids>314,776,780,1411,1427,27901,27902,45550,45551,46384,46808</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/25130858$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Kanazawa, Tomoko</creatorcontrib><creatorcontrib>Furumatsu, Takayuki</creatorcontrib><creatorcontrib>Matsumoto-Ogawa, Emi</creatorcontrib><creatorcontrib>Maehara, Ami</creatorcontrib><creatorcontrib>Ozaki, Toshifumi</creatorcontrib><title>Role of Rho small GTPases in meniscus cells</title><title>Journal of orthopaedic research</title><addtitle>J. Orthop. Res</addtitle><description>ABSTRACT
We previously reported that mechanical stretch regulates Sry‐type HMG box (SOX) 9‐dependent α1(II) collagen (COL2A1) expression in inner meniscus cells. This study examined the role of the small Rho guanosine 5' triphosphatase Rac1 and Rho‐associated kinase (ROCK) in the regulation of stretch‐induced SOX9 gene expression in cultured human inner meniscus cells. COL2A1 and SOX9 gene expression was assessed by real‐time PCR after application of uni‐axial cyclic tensile strain (CTS) in the presence or absence of ROCK and Rac1 inhibitors. The subcellular localization of SOX9 and the Rac1 effector cyclic AMP response element‐binding protein (CREB), the phosphorylation state of SOX9, Rac1 activation, and the binding of CREB to the SOX9 promoter were assessed. CTS increased the expression of COL2A1 and SOX9, which was suppressed by inhibition of Rac1. ROCK inhibition enhanced COL2A1 and SOX9 gene expression in the absence of CTS. CTS stimulated the nuclear translocation and phosphorylation of SOX9, and increased Rac1 activation. CTS also increased the binding of CREB to the SOX9 promoter. The results suggest that mechanical stretch‐dependent upregulation of SOX9 by CREB in inner meniscus cells depends on the antagonistic activities of ROCK and Rac1. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:1479–1486, 2014.</description><subject>Active Transport, Cell Nucleus</subject><subject>Animals</subject><subject>Arthroplasty, Replacement, Knee</subject><subject>Biomechanical Phenomena</subject><subject>Cartilage - enzymology</subject><subject>Collagen Type II - metabolism</subject><subject>CREB</subject><subject>Cyclic AMP Response Element-Binding Protein - metabolism</subject><subject>Gene Expression Regulation, Enzymologic</subject><subject>Humans</subject><subject>Knee - physiology</subject><subject>Mechanical stretch</subject><subject>Meniscus</subject><subject>Phosphorylation</subject><subject>Promoter Regions, Genetic</subject><subject>Rac1</subject><subject>rac1 GTP-Binding Protein - metabolism</subject><subject>Rats</subject><subject>rho GTP-Binding Proteins - metabolism</subject><subject>ROCK</subject><subject>SOX9</subject><subject>SOX9 Transcription Factor - metabolism</subject><subject>Stress, Mechanical</subject><subject>Tensile Strength</subject><issn>0736-0266</issn><issn>1554-527X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2014</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp1kM1OAjEURhujEUQXvoCZpcYMtLd_dKlERwkRQxDdNWVo4-AMg1OI8vZWB9i5uptzT04-hM4JbhOMoTMvqzaAxPQANQnnLOYg3w5RE0sqYgxCNNCJ93OMsSTQPUYN4ITiLu820fWozG1Uumj0Xka-MHkeJeNn462PskVU2EXm07WPUpvn_hQdOZN7e7a9LfRyfzfuPcSDYfLYuxnEKROMxk65KXcciHIKiINpqliIclwCF44I5RiVAA6bWUiw3EriBDAhATMDgtAWuqy9y6r8XFu_0kWoCAVmYcu114QLQQhnSgX0qkbTqvS-sk4vq6ww1UYTrH-30WEb_bdNYC-22vW0sLM9uRsjAJ0a-Mpyu_nfpPvD0U4Z1x-ZX9nv_YepPrSQVHL9-pTo8SSZKN7v6Vv6A0WjeZA</recordid><startdate>201411</startdate><enddate>201411</enddate><creator>Kanazawa, Tomoko</creator><creator>Furumatsu, Takayuki</creator><creator>Matsumoto-Ogawa, Emi</creator><creator>Maehara, Ami</creator><creator>Ozaki, Toshifumi</creator><general>Blackwell Publishing Ltd</general><scope>BSCLL</scope><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>201411</creationdate><title>Role of Rho small GTPases in meniscus cells</title><author>Kanazawa, Tomoko ; Furumatsu, Takayuki ; Matsumoto-Ogawa, Emi ; Maehara, Ami ; Ozaki, Toshifumi</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4643-f9fb5f5219f921f2bc94554f57256f169f43722f0ad858e5e71f62467204a2613</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2014</creationdate><topic>Active Transport, Cell Nucleus</topic><topic>Animals</topic><topic>Arthroplasty, Replacement, Knee</topic><topic>Biomechanical Phenomena</topic><topic>Cartilage - enzymology</topic><topic>Collagen Type II - metabolism</topic><topic>CREB</topic><topic>Cyclic AMP Response Element-Binding Protein - metabolism</topic><topic>Gene Expression Regulation, Enzymologic</topic><topic>Humans</topic><topic>Knee - physiology</topic><topic>Mechanical stretch</topic><topic>Meniscus</topic><topic>Phosphorylation</topic><topic>Promoter Regions, Genetic</topic><topic>Rac1</topic><topic>rac1 GTP-Binding Protein - metabolism</topic><topic>Rats</topic><topic>rho GTP-Binding Proteins - metabolism</topic><topic>ROCK</topic><topic>SOX9</topic><topic>SOX9 Transcription Factor - metabolism</topic><topic>Stress, Mechanical</topic><topic>Tensile Strength</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Kanazawa, Tomoko</creatorcontrib><creatorcontrib>Furumatsu, Takayuki</creatorcontrib><creatorcontrib>Matsumoto-Ogawa, Emi</creatorcontrib><creatorcontrib>Maehara, Ami</creatorcontrib><creatorcontrib>Ozaki, Toshifumi</creatorcontrib><collection>Istex</collection><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>Journal of orthopaedic research</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Kanazawa, Tomoko</au><au>Furumatsu, Takayuki</au><au>Matsumoto-Ogawa, Emi</au><au>Maehara, Ami</au><au>Ozaki, Toshifumi</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Role of Rho small GTPases in meniscus cells</atitle><jtitle>Journal of orthopaedic research</jtitle><addtitle>J. Orthop. Res</addtitle><date>2014-11</date><risdate>2014</risdate><volume>32</volume><issue>11</issue><spage>1479</spage><epage>1486</epage><pages>1479-1486</pages><issn>0736-0266</issn><eissn>1554-527X</eissn><abstract>ABSTRACT
We previously reported that mechanical stretch regulates Sry‐type HMG box (SOX) 9‐dependent α1(II) collagen (COL2A1) expression in inner meniscus cells. This study examined the role of the small Rho guanosine 5' triphosphatase Rac1 and Rho‐associated kinase (ROCK) in the regulation of stretch‐induced SOX9 gene expression in cultured human inner meniscus cells. COL2A1 and SOX9 gene expression was assessed by real‐time PCR after application of uni‐axial cyclic tensile strain (CTS) in the presence or absence of ROCK and Rac1 inhibitors. The subcellular localization of SOX9 and the Rac1 effector cyclic AMP response element‐binding protein (CREB), the phosphorylation state of SOX9, Rac1 activation, and the binding of CREB to the SOX9 promoter were assessed. CTS increased the expression of COL2A1 and SOX9, which was suppressed by inhibition of Rac1. ROCK inhibition enhanced COL2A1 and SOX9 gene expression in the absence of CTS. CTS stimulated the nuclear translocation and phosphorylation of SOX9, and increased Rac1 activation. CTS also increased the binding of CREB to the SOX9 promoter. The results suggest that mechanical stretch‐dependent upregulation of SOX9 by CREB in inner meniscus cells depends on the antagonistic activities of ROCK and Rac1. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:1479–1486, 2014.</abstract><cop>United States</cop><pub>Blackwell Publishing Ltd</pub><pmid>25130858</pmid><doi>10.1002/jor.22703</doi><tpages>8</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Active Transport, Cell Nucleus Animals Arthroplasty, Replacement, Knee Biomechanical Phenomena Cartilage - enzymology Collagen Type II - metabolism CREB Cyclic AMP Response Element-Binding Protein - metabolism Gene Expression Regulation, Enzymologic Humans Knee - physiology Mechanical stretch Meniscus Phosphorylation Promoter Regions, Genetic Rac1 rac1 GTP-Binding Protein - metabolism Rats rho GTP-Binding Proteins - metabolism ROCK SOX9 SOX9 Transcription Factor - metabolism Stress, Mechanical Tensile Strength |
| title | Role of Rho small GTPases in meniscus cells |
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