Lipopolysaccharide Induces the Migration of Human Dental Pulp Cells by Up-regulating miR-146a

Abstract Introduction MicroRNAs are small noncoding RNAs that play crucial roles in regulating normal and pathologic functions. Bacterial lipopolysaccharide (LPS) is one of the key regulators of pulpal pathogenesis. This study investigated how LPS regulates microRNA expression and affects the phenot...

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Veröffentlicht in:Journal of endodontics 2012-12, Vol.38 (12), p.1598-1603
Hauptverfasser: Wang, Min-Ching, DDS, Hung, Pei-Shih, PhD, Tu, Hsi-Feng, DDS, PhD, Shih, Wen-Yu, DDS, MS, Li, Wan-Chun, PhD, Chang, Kuo-Wei, DDS, PhD
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container_end_page 1603
container_issue 12
container_start_page 1598
container_title Journal of endodontics
container_volume 38
creator Wang, Min-Ching, DDS
Hung, Pei-Shih, PhD
Tu, Hsi-Feng, DDS, PhD
Shih, Wen-Yu, DDS, MS
Li, Wan-Chun, PhD
Chang, Kuo-Wei, DDS, PhD
description Abstract Introduction MicroRNAs are small noncoding RNAs that play crucial roles in regulating normal and pathologic functions. Bacterial lipopolysaccharide (LPS) is one of the key regulators of pulpal pathogenesis. This study investigated how LPS regulates microRNA expression and affects the phenotype of human dental pulp cells (DPCs). Methods Primary DPCs were established and immortalized to achieve immortalized DPCs (I-DPCs). DPCs and I-DPCs were treated with LPS and examined to identify changes in microRNA expression, cell proliferation, and cell migration. Quantitative reverse-transcriptase polymerase chain reaction was used to detect changes in gene expression. Exogenous miR-146a expression was performed transfection with pre -mir-146a mimic. Knockdown of interleukin receptor–associated kinase (IRAK1) and tumor necrosis factor receptor–associated factor 6 (TRAF6) expression was performed by small interference oligonucleotide transfection. Western blot analysis was used to detect changes in the expression of the IRAK1 and TRAF6 proteins. Results The differentiation of DPCs was induced by osteogenic medium. I-DPCs had a higher level of human telomerase reverse transcriptase gene than the parental DPCs. Up-regulation of miR-146a expression and an increase in migration was induced by LPS treatment of DPCs and I-DPCs. Exogenous miR-146a expression increased the migration of DPCs and I-DPCs and down-regulated the expression of IRAK1 and TRAF6. Knockdown of IRAK1 and/or TRAF6 increased the migration of DPCs. Conclusions The results suggested that LPS is able to increase the migration of DPCs by modulating the miR-146a –TRAF6/IRAK1 regulatory cascade.
doi_str_mv 10.1016/j.joen.2012.09.008
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Bacterial lipopolysaccharide (LPS) is one of the key regulators of pulpal pathogenesis. This study investigated how LPS regulates microRNA expression and affects the phenotype of human dental pulp cells (DPCs). Methods Primary DPCs were established and immortalized to achieve immortalized DPCs (I-DPCs). DPCs and I-DPCs were treated with LPS and examined to identify changes in microRNA expression, cell proliferation, and cell migration. Quantitative reverse-transcriptase polymerase chain reaction was used to detect changes in gene expression. Exogenous miR-146a expression was performed transfection with pre -mir-146a mimic. Knockdown of interleukin receptor–associated kinase (IRAK1) and tumor necrosis factor receptor–associated factor 6 (TRAF6) expression was performed by small interference oligonucleotide transfection. Western blot analysis was used to detect changes in the expression of the IRAK1 and TRAF6 proteins. Results The differentiation of DPCs was induced by osteogenic medium. I-DPCs had a higher level of human telomerase reverse transcriptase gene than the parental DPCs. Up-regulation of miR-146a expression and an increase in migration was induced by LPS treatment of DPCs and I-DPCs. Exogenous miR-146a expression increased the migration of DPCs and I-DPCs and down-regulated the expression of IRAK1 and TRAF6. Knockdown of IRAK1 and/or TRAF6 increased the migration of DPCs. Conclusions The results suggested that LPS is able to increase the migration of DPCs by modulating the miR-146a –TRAF6/IRAK1 regulatory cascade.</description><identifier>ISSN: 0099-2399</identifier><identifier>EISSN: 1878-3554</identifier><identifier>DOI: 10.1016/j.joen.2012.09.008</identifier><identifier>PMID: 23146644</identifier><language>eng</language><publisher>United States: Elsevier Inc</publisher><subject>Cell Culture Techniques ; Cell Differentiation - physiology ; Cell Line ; Cell Movement - drug effects ; Cell Proliferation - drug effects ; Culture Media ; Dental Pulp - cytology ; Dental Pulp - drug effects ; Dentistry ; Endocrinology &amp; Metabolism ; Gene Expression Regulation - drug effects ; Gene Knockdown Techniques ; Gene Silencing ; Humans ; Interleukin-1 Receptor-Associated Kinases - analysis ; Interleukin-1 Receptor-Associated Kinases - genetics ; IRAK1 ; Lipopolysaccharides - pharmacology ; LPS ; MicroRNAs - drug effects ; MicroRNAs - genetics ; migration ; miR-146a ; Osteogenesis - physiology ; Phenotype ; pulp ; Reverse Transcriptase Polymerase Chain Reaction ; RNA, Small Interfering - genetics ; TNF Receptor-Associated Factor 6 - analysis ; TNF Receptor-Associated Factor 6 - genetics ; TRAF6 ; Transfection ; Up-Regulation - drug effects</subject><ispartof>Journal of endodontics, 2012-12, Vol.38 (12), p.1598-1603</ispartof><rights>American Association of Endodontists</rights><rights>2012 American Association of Endodontists</rights><rights>Copyright © 2012 American Association of Endodontists. All rights reserved.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c510t-ad4922bc7b8db6a4e685ec5d500f3c53ac4ac7744de6c38a8a06bd3963fa433</citedby><cites>FETCH-LOGICAL-c510t-ad4922bc7b8db6a4e685ec5d500f3c53ac4ac7744de6c38a8a06bd3963fa433</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktohtml>$$Uhttps://dx.doi.org/10.1016/j.joen.2012.09.008$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,778,782,3539,27911,27912,45982</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/23146644$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wang, Min-Ching, DDS</creatorcontrib><creatorcontrib>Hung, Pei-Shih, PhD</creatorcontrib><creatorcontrib>Tu, Hsi-Feng, DDS, PhD</creatorcontrib><creatorcontrib>Shih, Wen-Yu, DDS, MS</creatorcontrib><creatorcontrib>Li, Wan-Chun, PhD</creatorcontrib><creatorcontrib>Chang, Kuo-Wei, DDS, PhD</creatorcontrib><title>Lipopolysaccharide Induces the Migration of Human Dental Pulp Cells by Up-regulating miR-146a</title><title>Journal of endodontics</title><addtitle>J Endod</addtitle><description>Abstract Introduction MicroRNAs are small noncoding RNAs that play crucial roles in regulating normal and pathologic functions. Bacterial lipopolysaccharide (LPS) is one of the key regulators of pulpal pathogenesis. This study investigated how LPS regulates microRNA expression and affects the phenotype of human dental pulp cells (DPCs). Methods Primary DPCs were established and immortalized to achieve immortalized DPCs (I-DPCs). DPCs and I-DPCs were treated with LPS and examined to identify changes in microRNA expression, cell proliferation, and cell migration. Quantitative reverse-transcriptase polymerase chain reaction was used to detect changes in gene expression. Exogenous miR-146a expression was performed transfection with pre -mir-146a mimic. Knockdown of interleukin receptor–associated kinase (IRAK1) and tumor necrosis factor receptor–associated factor 6 (TRAF6) expression was performed by small interference oligonucleotide transfection. Western blot analysis was used to detect changes in the expression of the IRAK1 and TRAF6 proteins. Results The differentiation of DPCs was induced by osteogenic medium. I-DPCs had a higher level of human telomerase reverse transcriptase gene than the parental DPCs. Up-regulation of miR-146a expression and an increase in migration was induced by LPS treatment of DPCs and I-DPCs. Exogenous miR-146a expression increased the migration of DPCs and I-DPCs and down-regulated the expression of IRAK1 and TRAF6. Knockdown of IRAK1 and/or TRAF6 increased the migration of DPCs. Conclusions The results suggested that LPS is able to increase the migration of DPCs by modulating the miR-146a –TRAF6/IRAK1 regulatory cascade.</description><subject>Cell Culture Techniques</subject><subject>Cell Differentiation - physiology</subject><subject>Cell Line</subject><subject>Cell Movement - drug effects</subject><subject>Cell Proliferation - drug effects</subject><subject>Culture Media</subject><subject>Dental Pulp - cytology</subject><subject>Dental Pulp - drug effects</subject><subject>Dentistry</subject><subject>Endocrinology &amp; Metabolism</subject><subject>Gene Expression Regulation - drug effects</subject><subject>Gene Knockdown Techniques</subject><subject>Gene Silencing</subject><subject>Humans</subject><subject>Interleukin-1 Receptor-Associated Kinases - analysis</subject><subject>Interleukin-1 Receptor-Associated Kinases - genetics</subject><subject>IRAK1</subject><subject>Lipopolysaccharides - pharmacology</subject><subject>LPS</subject><subject>MicroRNAs - drug effects</subject><subject>MicroRNAs - genetics</subject><subject>migration</subject><subject>miR-146a</subject><subject>Osteogenesis - physiology</subject><subject>Phenotype</subject><subject>pulp</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>RNA, Small Interfering - genetics</subject><subject>TNF Receptor-Associated Factor 6 - analysis</subject><subject>TNF Receptor-Associated Factor 6 - genetics</subject><subject>TRAF6</subject><subject>Transfection</subject><subject>Up-Regulation - drug effects</subject><issn>0099-2399</issn><issn>1878-3554</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2012</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNp9kU2L1TAUhoMoznX0D7iQLN205qtpCyLI9WMGriiOLiWkyemd1DbpJK1w_70pd3ThwlU2z_uek-cg9JySkhIqXw3lEMCXjFBWkrYkpHmAdrSpm4JXlXiIdoS0bcF4216gJykNhNCa8_oxumCcCimF2KEfBzeHOYynpI251dFZwNfergYSXm4Bf3LHqBcXPA49vlon7fE78Ise8Zd1nPEexjHh7oS_z0WE4zpm1h_x5L4WeYJ-ih71ekzw7P69RDcf3n_bXxWHzx-v928PhakoWQptRctYZ-qusZ3UAmRTgalsRUjPTcW1EdrUtRAWpOGNbjSRneWt5L0WnF-il-fWOYa7FdKiJpdM3kx7CGtSlFa0ZRWlMqPsjJoYUorQqzm6SceTokRtUtWgNqlqk6pIq7LUHHpx3792E9i_kT8WM_D6DED-4y8HUSXjwBuwLoJZlA3u__1v_omb0Xln9PgTTpCGsEaf7SmqUs6om-2s21Upy-laSP4bPoWdIw</recordid><startdate>20121201</startdate><enddate>20121201</enddate><creator>Wang, Min-Ching, DDS</creator><creator>Hung, Pei-Shih, PhD</creator><creator>Tu, Hsi-Feng, DDS, PhD</creator><creator>Shih, Wen-Yu, DDS, MS</creator><creator>Li, Wan-Chun, PhD</creator><creator>Chang, Kuo-Wei, DDS, PhD</creator><general>Elsevier Inc</general><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>20121201</creationdate><title>Lipopolysaccharide Induces the Migration of Human Dental Pulp Cells by Up-regulating miR-146a</title><author>Wang, Min-Ching, DDS ; Hung, Pei-Shih, PhD ; Tu, Hsi-Feng, DDS, PhD ; Shih, Wen-Yu, DDS, MS ; Li, Wan-Chun, PhD ; Chang, Kuo-Wei, DDS, PhD</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c510t-ad4922bc7b8db6a4e685ec5d500f3c53ac4ac7744de6c38a8a06bd3963fa433</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2012</creationdate><topic>Cell Culture Techniques</topic><topic>Cell Differentiation - physiology</topic><topic>Cell Line</topic><topic>Cell Movement - drug effects</topic><topic>Cell Proliferation - drug effects</topic><topic>Culture Media</topic><topic>Dental Pulp - cytology</topic><topic>Dental Pulp - drug effects</topic><topic>Dentistry</topic><topic>Endocrinology &amp; Metabolism</topic><topic>Gene Expression Regulation - drug effects</topic><topic>Gene Knockdown Techniques</topic><topic>Gene Silencing</topic><topic>Humans</topic><topic>Interleukin-1 Receptor-Associated Kinases - analysis</topic><topic>Interleukin-1 Receptor-Associated Kinases - genetics</topic><topic>IRAK1</topic><topic>Lipopolysaccharides - pharmacology</topic><topic>LPS</topic><topic>MicroRNAs - drug effects</topic><topic>MicroRNAs - genetics</topic><topic>migration</topic><topic>miR-146a</topic><topic>Osteogenesis - physiology</topic><topic>Phenotype</topic><topic>pulp</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>RNA, Small Interfering - genetics</topic><topic>TNF Receptor-Associated Factor 6 - analysis</topic><topic>TNF Receptor-Associated Factor 6 - genetics</topic><topic>TRAF6</topic><topic>Transfection</topic><topic>Up-Regulation - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Wang, Min-Ching, DDS</creatorcontrib><creatorcontrib>Hung, Pei-Shih, PhD</creatorcontrib><creatorcontrib>Tu, Hsi-Feng, DDS, PhD</creatorcontrib><creatorcontrib>Shih, Wen-Yu, DDS, MS</creatorcontrib><creatorcontrib>Li, Wan-Chun, PhD</creatorcontrib><creatorcontrib>Chang, Kuo-Wei, DDS, PhD</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>Journal of endodontics</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Wang, Min-Ching, DDS</au><au>Hung, Pei-Shih, PhD</au><au>Tu, Hsi-Feng, DDS, PhD</au><au>Shih, Wen-Yu, DDS, MS</au><au>Li, Wan-Chun, PhD</au><au>Chang, Kuo-Wei, DDS, PhD</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Lipopolysaccharide Induces the Migration of Human Dental Pulp Cells by Up-regulating miR-146a</atitle><jtitle>Journal of endodontics</jtitle><addtitle>J Endod</addtitle><date>2012-12-01</date><risdate>2012</risdate><volume>38</volume><issue>12</issue><spage>1598</spage><epage>1603</epage><pages>1598-1603</pages><issn>0099-2399</issn><eissn>1878-3554</eissn><abstract>Abstract Introduction MicroRNAs are small noncoding RNAs that play crucial roles in regulating normal and pathologic functions. Bacterial lipopolysaccharide (LPS) is one of the key regulators of pulpal pathogenesis. This study investigated how LPS regulates microRNA expression and affects the phenotype of human dental pulp cells (DPCs). Methods Primary DPCs were established and immortalized to achieve immortalized DPCs (I-DPCs). DPCs and I-DPCs were treated with LPS and examined to identify changes in microRNA expression, cell proliferation, and cell migration. Quantitative reverse-transcriptase polymerase chain reaction was used to detect changes in gene expression. Exogenous miR-146a expression was performed transfection with pre -mir-146a mimic. Knockdown of interleukin receptor–associated kinase (IRAK1) and tumor necrosis factor receptor–associated factor 6 (TRAF6) expression was performed by small interference oligonucleotide transfection. Western blot analysis was used to detect changes in the expression of the IRAK1 and TRAF6 proteins. Results The differentiation of DPCs was induced by osteogenic medium. I-DPCs had a higher level of human telomerase reverse transcriptase gene than the parental DPCs. Up-regulation of miR-146a expression and an increase in migration was induced by LPS treatment of DPCs and I-DPCs. Exogenous miR-146a expression increased the migration of DPCs and I-DPCs and down-regulated the expression of IRAK1 and TRAF6. Knockdown of IRAK1 and/or TRAF6 increased the migration of DPCs. Conclusions The results suggested that LPS is able to increase the migration of DPCs by modulating the miR-146a –TRAF6/IRAK1 regulatory cascade.</abstract><cop>United States</cop><pub>Elsevier Inc</pub><pmid>23146644</pmid><doi>10.1016/j.joen.2012.09.008</doi><tpages>6</tpages></addata></record>
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subjects Cell Culture Techniques
Cell Differentiation - physiology
Cell Line
Cell Movement - drug effects
Cell Proliferation - drug effects
Culture Media
Dental Pulp - cytology
Dental Pulp - drug effects
Dentistry
Endocrinology & Metabolism
Gene Expression Regulation - drug effects
Gene Knockdown Techniques
Gene Silencing
Humans
Interleukin-1 Receptor-Associated Kinases - analysis
Interleukin-1 Receptor-Associated Kinases - genetics
IRAK1
Lipopolysaccharides - pharmacology
LPS
MicroRNAs - drug effects
MicroRNAs - genetics
migration
miR-146a
Osteogenesis - physiology
Phenotype
pulp
Reverse Transcriptase Polymerase Chain Reaction
RNA, Small Interfering - genetics
TNF Receptor-Associated Factor 6 - analysis
TNF Receptor-Associated Factor 6 - genetics
TRAF6
Transfection
Up-Regulation - drug effects
title Lipopolysaccharide Induces the Migration of Human Dental Pulp Cells by Up-regulating miR-146a
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