Insulin‐like growth factor‐1 promotes osteogenic differentiation and collagen I alpha 2 synthesis via induction of mRNA‐binding protein LARP6 expression
This study explored the mechanism underlying the stimulation of collagen synthesis and osteoblastic differentiation by insulin‐like growth factor 1 (IGF1) in primary mouse osteoblasts. Primary mouse calvarial osteoblasts were cultured and treated with various doses of IGF1 before transfection with s...
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| Veröffentlicht in: | Development, growth & differentiation growth & differentiation, 2017-02, Vol.59 (2), p.94-103 |
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| creator | Guo, Yue Tang, Chen‐Yi Man, Xiao‐Fei Tang, Hao‐Neng Tang, Jun Zhou, Ci‐La Tan, Shu‐Wen Wang, Min Feng, Yun‐Zhi Zhou, Hou‐De |
| description | This study explored the mechanism underlying the stimulation of collagen synthesis and osteoblastic differentiation by insulin‐like growth factor 1 (IGF1) in primary mouse osteoblasts. Primary mouse calvarial osteoblasts were cultured and treated with various doses of IGF1 before transfection with siRNA targeting the collagen type I alpha 2 (Col1a2) or La ribonucleoprotein domain family member 6 (Larp6) genes. Alkaline phosphatase (ALP) activity, osteocalcin staining, alizarin red quantification and the expression level of runt‐related transcription factor 2 (RUNX2) were performed to assess the differentiation of pre‐osteoblasts. Based on Western blot analysis, IGF1 up‐regulated COL1A2 protein expression in the primary osteoblasts in a dose‐ and time‐dependent manner. In addition, Col1a2 interference inhibited the differentiation and mineralization of osteoblasts. IGF1 also stimulated the differentiation of mouse primary osteoblasts and increased LARP6 expression during osteogenic differentiation. RNA‐Immunoprecipitation (IP) indicated that LARP6 could bind to Col1a2 mRNA after IGF1 stimulation. However, transfection of Larp6‐specific siRNA significantly reduced collagen and ALP secretion, mineralization and inhibited the expression of osteocalcin and RUNX2, indicating that Larp6 interference inhibited the differentiation ability of primary mouse calvarial osteoblasts, and these effects could not be reversed by IGF1. Thus, IGF1 could promote COL1A2 expression and osteoblast differentiation in primary mouse calvarial pre‐osteoblasts by increasing LARP6 expression via a posttranscriptional mechanism.
IGF1 promotes primary osteoblast differentiation and Col1a2 expression. IGF1 induces LARP6 expression. LARP6 regulates Col1a2 expression and primary osteoblast differentiation. |
| doi_str_mv | 10.1111/dgd.12342 |
| format | Article |
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IGF1 promotes primary osteoblast differentiation and Col1a2 expression. IGF1 induces LARP6 expression. LARP6 regulates Col1a2 expression and primary osteoblast differentiation.</description><identifier>ISSN: 0012-1592</identifier><identifier>EISSN: 1440-169X</identifier><identifier>DOI: 10.1111/dgd.12342</identifier><identifier>PMID: 28211947</identifier><identifier>CODEN: DGDFA5</identifier><language>eng</language><publisher>Japan: Wiley Subscription Services, Inc</publisher><subject>Alkaline Phosphatase - metabolism ; Animals ; Animals, Newborn ; Autoantigens - genetics ; Autoantigens - metabolism ; Blotting, Western ; Cell Differentiation - drug effects ; Cell Differentiation - genetics ; Cells, Cultured ; COL1A2 ; Collagen ; Collagen Type I - biosynthesis ; Collagen Type I - genetics ; Core Binding Factor Alpha 1 Subunit - genetics ; Core Binding Factor Alpha 1 Subunit - metabolism ; Dose-Response Relationship, Drug ; Gene Expression - drug effects ; Genes ; Insulin-Like Growth Factor I - pharmacology ; Insulin-like growth factors ; insulin‐like growth factor 1 ; La ribonucleoprotein domain family member 6 ; Mice, Inbred C57BL ; Mineralization ; osteoblast ; Osteoblasts - cytology ; Osteoblasts - drug effects ; Osteoblasts - metabolism ; Osteocalcin - metabolism ; Osteogenesis - drug effects ; Osteogenesis - genetics ; osteogenic differentiation ; Protein Binding ; Reverse Transcriptase Polymerase Chain Reaction ; Ribonucleoproteins - genetics ; Ribonucleoproteins - metabolism ; RNA Interference ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; RNA-Binding Proteins - genetics ; RNA-Binding Proteins - metabolism ; SS-B Antigen ; Time Factors</subject><ispartof>Development, growth & differentiation, 2017-02, Vol.59 (2), p.94-103</ispartof><rights>2017 Japanese Society of Developmental Biologists</rights><rights>2017 Japanese Society of Developmental Biologists.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://onlinelibrary.wiley.com/doi/pdf/10.1111%2Fdgd.12342$$EPDF$$P50$$Gwiley$$H</linktopdf><linktohtml>$$Uhttps://onlinelibrary.wiley.com/doi/full/10.1111%2Fdgd.12342$$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/28211947$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Guo, Yue</creatorcontrib><creatorcontrib>Tang, Chen‐Yi</creatorcontrib><creatorcontrib>Man, Xiao‐Fei</creatorcontrib><creatorcontrib>Tang, Hao‐Neng</creatorcontrib><creatorcontrib>Tang, Jun</creatorcontrib><creatorcontrib>Zhou, Ci‐La</creatorcontrib><creatorcontrib>Tan, Shu‐Wen</creatorcontrib><creatorcontrib>Wang, Min</creatorcontrib><creatorcontrib>Feng, Yun‐Zhi</creatorcontrib><creatorcontrib>Zhou, Hou‐De</creatorcontrib><title>Insulin‐like growth factor‐1 promotes osteogenic differentiation and collagen I alpha 2 synthesis via induction of mRNA‐binding protein LARP6 expression</title><title>Development, growth & differentiation</title><addtitle>Dev Growth Differ</addtitle><description>This study explored the mechanism underlying the stimulation of collagen synthesis and osteoblastic differentiation by insulin‐like growth factor 1 (IGF1) in primary mouse osteoblasts. Primary mouse calvarial osteoblasts were cultured and treated with various doses of IGF1 before transfection with siRNA targeting the collagen type I alpha 2 (Col1a2) or La ribonucleoprotein domain family member 6 (Larp6) genes. Alkaline phosphatase (ALP) activity, osteocalcin staining, alizarin red quantification and the expression level of runt‐related transcription factor 2 (RUNX2) were performed to assess the differentiation of pre‐osteoblasts. Based on Western blot analysis, IGF1 up‐regulated COL1A2 protein expression in the primary osteoblasts in a dose‐ and time‐dependent manner. In addition, Col1a2 interference inhibited the differentiation and mineralization of osteoblasts. IGF1 also stimulated the differentiation of mouse primary osteoblasts and increased LARP6 expression during osteogenic differentiation. RNA‐Immunoprecipitation (IP) indicated that LARP6 could bind to Col1a2 mRNA after IGF1 stimulation. However, transfection of Larp6‐specific siRNA significantly reduced collagen and ALP secretion, mineralization and inhibited the expression of osteocalcin and RUNX2, indicating that Larp6 interference inhibited the differentiation ability of primary mouse calvarial osteoblasts, and these effects could not be reversed by IGF1. Thus, IGF1 could promote COL1A2 expression and osteoblast differentiation in primary mouse calvarial pre‐osteoblasts by increasing LARP6 expression via a posttranscriptional mechanism.
IGF1 promotes primary osteoblast differentiation and Col1a2 expression. IGF1 induces LARP6 expression. LARP6 regulates Col1a2 expression and primary osteoblast differentiation.</description><subject>Alkaline Phosphatase - metabolism</subject><subject>Animals</subject><subject>Animals, Newborn</subject><subject>Autoantigens - genetics</subject><subject>Autoantigens - metabolism</subject><subject>Blotting, Western</subject><subject>Cell Differentiation - drug effects</subject><subject>Cell Differentiation - genetics</subject><subject>Cells, Cultured</subject><subject>COL1A2</subject><subject>Collagen</subject><subject>Collagen Type I - biosynthesis</subject><subject>Collagen Type I - genetics</subject><subject>Core Binding Factor Alpha 1 Subunit - genetics</subject><subject>Core Binding Factor Alpha 1 Subunit - metabolism</subject><subject>Dose-Response Relationship, Drug</subject><subject>Gene Expression - drug effects</subject><subject>Genes</subject><subject>Insulin-Like Growth Factor I - pharmacology</subject><subject>Insulin-like growth factors</subject><subject>insulin‐like growth factor 1</subject><subject>La ribonucleoprotein domain family member 6</subject><subject>Mice, Inbred C57BL</subject><subject>Mineralization</subject><subject>osteoblast</subject><subject>Osteoblasts - cytology</subject><subject>Osteoblasts - drug effects</subject><subject>Osteoblasts - metabolism</subject><subject>Osteocalcin - metabolism</subject><subject>Osteogenesis - drug effects</subject><subject>Osteogenesis - genetics</subject><subject>osteogenic differentiation</subject><subject>Protein Binding</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Ribonucleoproteins - genetics</subject><subject>Ribonucleoproteins - metabolism</subject><subject>RNA Interference</subject><subject>RNA, Messenger - genetics</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA-Binding Proteins - genetics</subject><subject>RNA-Binding Proteins - metabolism</subject><subject>SS-B Antigen</subject><subject>Time Factors</subject><issn>0012-1592</issn><issn>1440-169X</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2017</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNo9kclOAzEMhiMEgrIceAEUifNAttmOVQulUgUIgcRtlMnSpkyTMslQeuMReAIejichlMUHO7I_2bF_AI4xOsPRzuVUnmFCGdkCPcwYSnBWPm6DHkKYJDgtyR7Y936OEGIMk12wRwqCccnyHvgYW981xn6-vTfmScFp61ZhBjUXwbUxieGydQsXlIfOB-WmyhoBpdFatcoGw4NxFnIroXBNw2MZjiFvljMOCfRrG2bKGw9fDIfGyk5scKfh4u66H9vXMWns9HtIUMbCSf_uNoPqddkq7yN6CHY0b7w6-o0H4OHy4n5wlUxuRuNBf5LMKS1IIjTPdC5zkRa4oISVeVxdqTrlEvHo67pGTHKtC4HKVLCMyJLoWlJeC5aijB6A05--8SPPnfKhmruutXFkhYucspJGF6mTX6qrF0pWy9YseLuu_s4ZgfMfYGUatf6vY1R961RFnaqNTtVwNNw86BdcUIrO</recordid><startdate>201702</startdate><enddate>201702</enddate><creator>Guo, Yue</creator><creator>Tang, Chen‐Yi</creator><creator>Man, Xiao‐Fei</creator><creator>Tang, Hao‐Neng</creator><creator>Tang, Jun</creator><creator>Zhou, Ci‐La</creator><creator>Tan, Shu‐Wen</creator><creator>Wang, Min</creator><creator>Feng, Yun‐Zhi</creator><creator>Zhou, Hou‐De</creator><general>Wiley Subscription Services, Inc</general><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>7SS</scope><scope>7TK</scope><scope>7TM</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>RC3</scope></search><sort><creationdate>201702</creationdate><title>Insulin‐like growth factor‐1 promotes osteogenic differentiation and collagen I alpha 2 synthesis via induction of mRNA‐binding protein LARP6 expression</title><author>Guo, Yue ; Tang, Chen‐Yi ; Man, Xiao‐Fei ; Tang, Hao‐Neng ; Tang, Jun ; Zhou, Ci‐La ; Tan, Shu‐Wen ; Wang, Min ; Feng, Yun‐Zhi ; Zhou, Hou‐De</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-j3382-cfa6f7d7c581832497001eeb5ad0ab5abbb04daff8c095c462d92fbd3abc45063</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2017</creationdate><topic>Alkaline Phosphatase - metabolism</topic><topic>Animals</topic><topic>Animals, Newborn</topic><topic>Autoantigens - genetics</topic><topic>Autoantigens - metabolism</topic><topic>Blotting, Western</topic><topic>Cell Differentiation - drug effects</topic><topic>Cell Differentiation - genetics</topic><topic>Cells, Cultured</topic><topic>COL1A2</topic><topic>Collagen</topic><topic>Collagen Type I - biosynthesis</topic><topic>Collagen Type I - genetics</topic><topic>Core Binding Factor Alpha 1 Subunit - genetics</topic><topic>Core Binding Factor Alpha 1 Subunit - metabolism</topic><topic>Dose-Response Relationship, Drug</topic><topic>Gene Expression - drug effects</topic><topic>Genes</topic><topic>Insulin-Like Growth Factor I - pharmacology</topic><topic>Insulin-like growth factors</topic><topic>insulin‐like growth factor 1</topic><topic>La ribonucleoprotein domain family member 6</topic><topic>Mice, Inbred C57BL</topic><topic>Mineralization</topic><topic>osteoblast</topic><topic>Osteoblasts - cytology</topic><topic>Osteoblasts - drug effects</topic><topic>Osteoblasts - metabolism</topic><topic>Osteocalcin - metabolism</topic><topic>Osteogenesis - drug effects</topic><topic>Osteogenesis - genetics</topic><topic>osteogenic differentiation</topic><topic>Protein Binding</topic><topic>Reverse Transcriptase Polymerase Chain Reaction</topic><topic>Ribonucleoproteins - genetics</topic><topic>Ribonucleoproteins - metabolism</topic><topic>RNA Interference</topic><topic>RNA, Messenger - genetics</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA-Binding Proteins - genetics</topic><topic>RNA-Binding Proteins - metabolism</topic><topic>SS-B Antigen</topic><topic>Time Factors</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Guo, Yue</creatorcontrib><creatorcontrib>Tang, Chen‐Yi</creatorcontrib><creatorcontrib>Man, Xiao‐Fei</creatorcontrib><creatorcontrib>Tang, Hao‐Neng</creatorcontrib><creatorcontrib>Tang, Jun</creatorcontrib><creatorcontrib>Zhou, Ci‐La</creatorcontrib><creatorcontrib>Tan, Shu‐Wen</creatorcontrib><creatorcontrib>Wang, Min</creatorcontrib><creatorcontrib>Feng, Yun‐Zhi</creatorcontrib><creatorcontrib>Zhou, Hou‐De</creatorcontrib><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>Entomology Abstracts (Full archive)</collection><collection>Neurosciences Abstracts</collection><collection>Nucleic Acids Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>Genetics Abstracts</collection><jtitle>Development, growth & differentiation</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Guo, Yue</au><au>Tang, Chen‐Yi</au><au>Man, Xiao‐Fei</au><au>Tang, Hao‐Neng</au><au>Tang, Jun</au><au>Zhou, Ci‐La</au><au>Tan, Shu‐Wen</au><au>Wang, Min</au><au>Feng, Yun‐Zhi</au><au>Zhou, Hou‐De</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Insulin‐like growth factor‐1 promotes osteogenic differentiation and collagen I alpha 2 synthesis via induction of mRNA‐binding protein LARP6 expression</atitle><jtitle>Development, growth & differentiation</jtitle><addtitle>Dev Growth Differ</addtitle><date>2017-02</date><risdate>2017</risdate><volume>59</volume><issue>2</issue><spage>94</spage><epage>103</epage><pages>94-103</pages><issn>0012-1592</issn><eissn>1440-169X</eissn><coden>DGDFA5</coden><abstract>This study explored the mechanism underlying the stimulation of collagen synthesis and osteoblastic differentiation by insulin‐like growth factor 1 (IGF1) in primary mouse osteoblasts. Primary mouse calvarial osteoblasts were cultured and treated with various doses of IGF1 before transfection with siRNA targeting the collagen type I alpha 2 (Col1a2) or La ribonucleoprotein domain family member 6 (Larp6) genes. Alkaline phosphatase (ALP) activity, osteocalcin staining, alizarin red quantification and the expression level of runt‐related transcription factor 2 (RUNX2) were performed to assess the differentiation of pre‐osteoblasts. Based on Western blot analysis, IGF1 up‐regulated COL1A2 protein expression in the primary osteoblasts in a dose‐ and time‐dependent manner. In addition, Col1a2 interference inhibited the differentiation and mineralization of osteoblasts. IGF1 also stimulated the differentiation of mouse primary osteoblasts and increased LARP6 expression during osteogenic differentiation. RNA‐Immunoprecipitation (IP) indicated that LARP6 could bind to Col1a2 mRNA after IGF1 stimulation. However, transfection of Larp6‐specific siRNA significantly reduced collagen and ALP secretion, mineralization and inhibited the expression of osteocalcin and RUNX2, indicating that Larp6 interference inhibited the differentiation ability of primary mouse calvarial osteoblasts, and these effects could not be reversed by IGF1. Thus, IGF1 could promote COL1A2 expression and osteoblast differentiation in primary mouse calvarial pre‐osteoblasts by increasing LARP6 expression via a posttranscriptional mechanism.
IGF1 promotes primary osteoblast differentiation and Col1a2 expression. IGF1 induces LARP6 expression. LARP6 regulates Col1a2 expression and primary osteoblast differentiation.</abstract><cop>Japan</cop><pub>Wiley Subscription Services, Inc</pub><pmid>28211947</pmid><doi>10.1111/dgd.12342</doi><tpages>10</tpages><oa>free_for_read</oa></addata></record> |
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| subjects | Alkaline Phosphatase - metabolism Animals Animals, Newborn Autoantigens - genetics Autoantigens - metabolism Blotting, Western Cell Differentiation - drug effects Cell Differentiation - genetics Cells, Cultured COL1A2 Collagen Collagen Type I - biosynthesis Collagen Type I - genetics Core Binding Factor Alpha 1 Subunit - genetics Core Binding Factor Alpha 1 Subunit - metabolism Dose-Response Relationship, Drug Gene Expression - drug effects Genes Insulin-Like Growth Factor I - pharmacology Insulin-like growth factors insulin‐like growth factor 1 La ribonucleoprotein domain family member 6 Mice, Inbred C57BL Mineralization osteoblast Osteoblasts - cytology Osteoblasts - drug effects Osteoblasts - metabolism Osteocalcin - metabolism Osteogenesis - drug effects Osteogenesis - genetics osteogenic differentiation Protein Binding Reverse Transcriptase Polymerase Chain Reaction Ribonucleoproteins - genetics Ribonucleoproteins - metabolism RNA Interference RNA, Messenger - genetics RNA, Messenger - metabolism RNA-Binding Proteins - genetics RNA-Binding Proteins - metabolism SS-B Antigen Time Factors |
| title | Insulin‐like growth factor‐1 promotes osteogenic differentiation and collagen I alpha 2 synthesis via induction of mRNA‐binding protein LARP6 expression |
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