Influence of MicroRNA‐141 on Inhibition of the Proliferation of Bone Marrow Mesenchymal Stem Cells in Steroid‐Induced Osteonecrosis via SOX11
Objective To investigate whether miR‐141 and the sex determination region of Y chromosome box 11 (SOX11) play roles in steroid‐induced avascular necrosis of the femoral head (SANFH), and to explore whether miR‐141 could target SOX11 to influence the proliferation of bone marrow mesenchymal stem cell...
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| creator | Meng, Chen‐yang Xue, Fei Zhao, Zhen‐qun Hao, Ting Guo, Shi‐bing Feng, Wei |
| description | Objective
To investigate whether miR‐141 and the sex determination region of Y chromosome box 11 (SOX11) play roles in steroid‐induced avascular necrosis of the femoral head (SANFH), and to explore whether miR‐141 could target SOX11 to influence the proliferation of bone marrow mesenchymal stem cells (BMSC).
Methods
Bone marrow mesenchymal stem cells (BMSC) were isolated and cultured from 4‐week‐old Sprague Dawley rats. A flow cytometry assay was performed to identify BMSC. BMSC were divided into two groups: a control group and a dexamethasone (DEX) group. BMSC were transfected by miR‐141 mimic, miR‐141 inhibitor, and SOX11. Real‐time polymerase chain reaction (PCR) assay was performed to investigate the mRNA expression of miR‐141 and SOX11. The results were used to determine the effect of transfection and to verify the expression in each group and the association between miR‐141 and SOX11. Luciferase reporter assay revealed the targeted binding site between miR‐141 and the 3′‐untranslated region of SOX11 mRNA. MTT assays were performed to investigate the proliferation of BMSC in the miR‐141 mimic, miR‐141 inhibitor, and SOX11 groups.
Result
The results of the flow cytometry assay suggested that cells were positive for CD29 and CD90 while negative for CD45. This meant that the isolated and cultured cells were not hematopoietic stem cells. In addition, cell transfection was successful based on the expression of miR‐141 and SOX11. According to the results of real‐time PCR assay, the mRNA expression of miR‐141 in SANFH was upregulated (4.117 ± 0.042 vs 1 ± 0.027, P |
| doi_str_mv | 10.1111/os.12603 |
| format | Article |
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To investigate whether miR‐141 and the sex determination region of Y chromosome box 11 (SOX11) play roles in steroid‐induced avascular necrosis of the femoral head (SANFH), and to explore whether miR‐141 could target SOX11 to influence the proliferation of bone marrow mesenchymal stem cells (BMSC).
Methods
Bone marrow mesenchymal stem cells (BMSC) were isolated and cultured from 4‐week‐old Sprague Dawley rats. A flow cytometry assay was performed to identify BMSC. BMSC were divided into two groups: a control group and a dexamethasone (DEX) group. BMSC were transfected by miR‐141 mimic, miR‐141 inhibitor, and SOX11. Real‐time polymerase chain reaction (PCR) assay was performed to investigate the mRNA expression of miR‐141 and SOX11. The results were used to determine the effect of transfection and to verify the expression in each group and the association between miR‐141 and SOX11. Luciferase reporter assay revealed the targeted binding site between miR‐141 and the 3′‐untranslated region of SOX11 mRNA. MTT assays were performed to investigate the proliferation of BMSC in the miR‐141 mimic, miR‐141 inhibitor, and SOX11 groups.
Result
The results of the flow cytometry assay suggested that cells were positive for CD29 and CD90 while negative for CD45. This meant that the isolated and cultured cells were not hematopoietic stem cells. In addition, cell transfection was successful based on the expression of miR‐141 and SOX11. According to the results of real‐time PCR assay, the mRNA expression of miR‐141 in SANFH was upregulated (4.117 ± 0.042 vs 1 ± 0.027, P < 0.001), while SOX11 was downregulated (0.611 ± 0.055 vs 1 ± 0.027, P < 0.001) compared with the control group. Based on the results of the luciferase experiment, MiR‐141 could directly target the expression of SOX11. Inhibition of miR‐141 could upregulate the expression of SOX11 (2.623 ± 0.220 vs 1 ± 0.095, P < 0.001) according to the results of a real‐time PCR assay. MiR‐141 inhibited the proliferation of BMSC (0.618 ± 0.092 vs 1.004 ± 0.082, P < 0.001), while suppression of miR‐141 increased the proliferation of BMSC (0.960 ± 0.095 vs 0.742 ± 0.091, P < 0.001). Furthermore, according to the results of the MTT assay, SOX11 promoted the proliferation of BMSC (1.064 ± 0.093 vs 0.747 ± 0.090, P < 0.001).
Conclusion
MiR‐141 inhibited the proliferation of BMSC in SANFH by targeting SOX11. Inhibition of miR‐141 upregulated the expression of SOX11 and promoted the proliferation of BMSC. MiR‐141 and SOX11 could be new targets for investigating the mechanism of SANFH.]]></description><identifier>ISSN: 1757-7853</identifier><identifier>EISSN: 1757-7861</identifier><identifier>DOI: 10.1111/os.12603</identifier><identifier>PMID: 31916393</identifier><language>eng</language><publisher>Melbourne: John Wiley & Sons Australia, Ltd</publisher><subject>Adipocytes ; Bone marrow ; Bone marrow mesenchymal stem cells ; Cell culture ; Cell growth ; Flow cytometry ; Gene expression ; Glucocorticoid ; Life Sciences & Biomedicine ; Lipids ; Metabolism ; MicroRNAs ; MicroRNA‐141 ; Monoclonal antibodies ; Orthopedics ; Osteoporosis ; Polymerase chain reaction ; Science & Technology ; Scientific ; SOX11 ; Stem cells ; Steroid induced avascular necrosis of femoral head ; Y chromosomes</subject><ispartof>Orthopaedic surgery, 2020-02, Vol.12 (1), p.277-285</ispartof><rights>2020 The Authors. Orthopaedic Surgery published by Chinese Orthopaedic Association and John Wiley & Sons Australia, Ltd.</rights><rights>2020. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>true</woscitedreferencessubscribed><woscitedreferencescount>14</woscitedreferencescount><woscitedreferencesoriginalsourcerecordid>wos000506129800001</woscitedreferencesoriginalsourcerecordid><citedby>FETCH-LOGICAL-c5323-67a908afebec97472433660a0196fb8f370a90e2cea94be7cbcbfa07bf1250123</citedby><cites>FETCH-LOGICAL-c5323-67a908afebec97472433660a0196fb8f370a90e2cea94be7cbcbfa07bf1250123</cites><orcidid>0000-0001-7913-2011</orcidid></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7031553/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC7031553/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,728,781,785,865,886,1418,2103,2115,11567,27929,27930,28253,45579,45580,46057,46481,53796,53798</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/31916393$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Meng, Chen‐yang</creatorcontrib><creatorcontrib>Xue, Fei</creatorcontrib><creatorcontrib>Zhao, Zhen‐qun</creatorcontrib><creatorcontrib>Hao, Ting</creatorcontrib><creatorcontrib>Guo, Shi‐bing</creatorcontrib><creatorcontrib>Feng, Wei</creatorcontrib><title>Influence of MicroRNA‐141 on Inhibition of the Proliferation of Bone Marrow Mesenchymal Stem Cells in Steroid‐Induced Osteonecrosis via SOX11</title><title>Orthopaedic surgery</title><addtitle>ORTHOP SURG</addtitle><addtitle>Orthop Surg</addtitle><description><![CDATA[Objective
To investigate whether miR‐141 and the sex determination region of Y chromosome box 11 (SOX11) play roles in steroid‐induced avascular necrosis of the femoral head (SANFH), and to explore whether miR‐141 could target SOX11 to influence the proliferation of bone marrow mesenchymal stem cells (BMSC).
Methods
Bone marrow mesenchymal stem cells (BMSC) were isolated and cultured from 4‐week‐old Sprague Dawley rats. A flow cytometry assay was performed to identify BMSC. BMSC were divided into two groups: a control group and a dexamethasone (DEX) group. BMSC were transfected by miR‐141 mimic, miR‐141 inhibitor, and SOX11. Real‐time polymerase chain reaction (PCR) assay was performed to investigate the mRNA expression of miR‐141 and SOX11. The results were used to determine the effect of transfection and to verify the expression in each group and the association between miR‐141 and SOX11. Luciferase reporter assay revealed the targeted binding site between miR‐141 and the 3′‐untranslated region of SOX11 mRNA. MTT assays were performed to investigate the proliferation of BMSC in the miR‐141 mimic, miR‐141 inhibitor, and SOX11 groups.
Result
The results of the flow cytometry assay suggested that cells were positive for CD29 and CD90 while negative for CD45. This meant that the isolated and cultured cells were not hematopoietic stem cells. In addition, cell transfection was successful based on the expression of miR‐141 and SOX11. According to the results of real‐time PCR assay, the mRNA expression of miR‐141 in SANFH was upregulated (4.117 ± 0.042 vs 1 ± 0.027, P < 0.001), while SOX11 was downregulated (0.611 ± 0.055 vs 1 ± 0.027, P < 0.001) compared with the control group. Based on the results of the luciferase experiment, MiR‐141 could directly target the expression of SOX11. Inhibition of miR‐141 could upregulate the expression of SOX11 (2.623 ± 0.220 vs 1 ± 0.095, P < 0.001) according to the results of a real‐time PCR assay. MiR‐141 inhibited the proliferation of BMSC (0.618 ± 0.092 vs 1.004 ± 0.082, P < 0.001), while suppression of miR‐141 increased the proliferation of BMSC (0.960 ± 0.095 vs 0.742 ± 0.091, P < 0.001). Furthermore, according to the results of the MTT assay, SOX11 promoted the proliferation of BMSC (1.064 ± 0.093 vs 0.747 ± 0.090, P < 0.001).
Conclusion
MiR‐141 inhibited the proliferation of BMSC in SANFH by targeting SOX11. Inhibition of miR‐141 upregulated the expression of SOX11 and promoted the proliferation of BMSC. MiR‐141 and SOX11 could be new targets for investigating the mechanism of SANFH.]]></description><subject>Adipocytes</subject><subject>Bone marrow</subject><subject>Bone marrow mesenchymal stem cells</subject><subject>Cell culture</subject><subject>Cell growth</subject><subject>Flow cytometry</subject><subject>Gene expression</subject><subject>Glucocorticoid</subject><subject>Life Sciences & Biomedicine</subject><subject>Lipids</subject><subject>Metabolism</subject><subject>MicroRNAs</subject><subject>MicroRNA‐141</subject><subject>Monoclonal antibodies</subject><subject>Orthopedics</subject><subject>Osteoporosis</subject><subject>Polymerase chain reaction</subject><subject>Science & Technology</subject><subject>Scientific</subject><subject>SOX11</subject><subject>Stem cells</subject><subject>Steroid induced avascular necrosis of femoral head</subject><subject>Y chromosomes</subject><issn>1757-7853</issn><issn>1757-7861</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>24P</sourceid><sourceid>WIN</sourceid><sourceid>AOWDO</sourceid><sourceid>ABUWG</sourceid><sourceid>AFKRA</sourceid><sourceid>AZQEC</sourceid><sourceid>BENPR</sourceid><sourceid>CCPQU</sourceid><sourceid>DWQXO</sourceid><sourceid>DOA</sourceid><recordid>eNqNks1u1DAUhSMEoqUg8QTIEhskNMU3Tuxkg1RG_IzUYRADEjvLdq47HmXi1k5azY5HgFfkSfD8dEQXILzJ9fV3T07ik2VPgZ5CWq98PIWcU3YvOwZRipGoONw_1CU7yh7FuKSU10yIh9kRgxo4q9lx9nPS2XbAziDxlkydCf7zx7Nf339AAcR3ZNItnHa9S2U67xdIPgXfOotB3Tbf-A7JVIXgb8gUY9JarFeqJfMeV2SMbRuJ6za74F2TlCddMxhsyCz2mEbTG6OL5NopMp99A3icPbCqjfhk_zzJvr57-2X8YXQ-ez8Zn52PTMlyNuJC1bRSFjWaWhQiLxjjnCoKNbe6skzQBGBuUNWFRmG00VZRoS3kJYWcnWSTnW7j1VJeBrdSYS29cnLb8OFCqtA706IsqciprgQvAQqdNxqQK2FAiEI3VdEkrdc7rctBr7Ax2PVBtXdE7550biEv_LUUlEFZsiTwfC8Q_NWAsZdLP4Qufb_Mi0rQukpX90-KlRWwMhlM1IsdtfmzMaA9-AAqN3GRPsptXBL67E_fB_A2HwmodsANam-jcZukHDBKaUk55HWVKgpj129DMfZD16fRl_8_mujRnnYtrv_qWM7mO-e_ARSt6q8</recordid><startdate>202002</startdate><enddate>202002</enddate><creator>Meng, Chen‐yang</creator><creator>Xue, Fei</creator><creator>Zhao, Zhen‐qun</creator><creator>Hao, Ting</creator><creator>Guo, Shi‐bing</creator><creator>Feng, Wei</creator><general>John Wiley & Sons Australia, Ltd</general><general>Wiley</general><general>John Wiley & Sons, Inc</general><scope>24P</scope><scope>WIN</scope><scope>AOWDO</scope><scope>BLEPL</scope><scope>DTL</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>3V.</scope><scope>7X7</scope><scope>7XB</scope><scope>8FI</scope><scope>8FJ</scope><scope>8FK</scope><scope>ABUWG</scope><scope>AFKRA</scope><scope>AZQEC</scope><scope>BENPR</scope><scope>CCPQU</scope><scope>DWQXO</scope><scope>FYUFA</scope><scope>GHDGH</scope><scope>K9.</scope><scope>M0S</scope><scope>PIMPY</scope><scope>PQEST</scope><scope>PQQKQ</scope><scope>PQUKI</scope><scope>PRINS</scope><scope>5PM</scope><scope>DOA</scope><orcidid>https://orcid.org/0000-0001-7913-2011</orcidid></search><sort><creationdate>202002</creationdate><title>Influence of MicroRNA‐141 on Inhibition of the Proliferation of Bone Marrow Mesenchymal Stem Cells in Steroid‐Induced Osteonecrosis via SOX11</title><author>Meng, Chen‐yang ; Xue, Fei ; Zhao, Zhen‐qun ; Hao, Ting ; Guo, Shi‐bing ; Feng, Wei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5323-67a908afebec97472433660a0196fb8f370a90e2cea94be7cbcbfa07bf1250123</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2020</creationdate><topic>Adipocytes</topic><topic>Bone marrow</topic><topic>Bone marrow mesenchymal stem cells</topic><topic>Cell culture</topic><topic>Cell growth</topic><topic>Flow cytometry</topic><topic>Gene expression</topic><topic>Glucocorticoid</topic><topic>Life Sciences & Biomedicine</topic><topic>Lipids</topic><topic>Metabolism</topic><topic>MicroRNAs</topic><topic>MicroRNA‐141</topic><topic>Monoclonal antibodies</topic><topic>Orthopedics</topic><topic>Osteoporosis</topic><topic>Polymerase chain reaction</topic><topic>Science & Technology</topic><topic>Scientific</topic><topic>SOX11</topic><topic>Stem cells</topic><topic>Steroid induced avascular necrosis of femoral head</topic><topic>Y chromosomes</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Meng, Chen‐yang</creatorcontrib><creatorcontrib>Xue, Fei</creatorcontrib><creatorcontrib>Zhao, Zhen‐qun</creatorcontrib><creatorcontrib>Hao, Ting</creatorcontrib><creatorcontrib>Guo, Shi‐bing</creatorcontrib><creatorcontrib>Feng, Wei</creatorcontrib><collection>Wiley-Blackwell Open Access Titles</collection><collection>Wiley Free Content</collection><collection>Web of Science - Science Citation Index Expanded - 2020</collection><collection>Web of Science Core Collection</collection><collection>Science Citation Index Expanded</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>ProQuest Central (Corporate)</collection><collection>Health & Medical Collection</collection><collection>ProQuest Central (purchase pre-March 2016)</collection><collection>Hospital Premium Collection</collection><collection>Hospital Premium Collection (Alumni Edition)</collection><collection>ProQuest Central (Alumni) (purchase pre-March 2016)</collection><collection>ProQuest Central (Alumni Edition)</collection><collection>ProQuest Central UK/Ireland</collection><collection>ProQuest Central Essentials</collection><collection>ProQuest Central</collection><collection>ProQuest One Community College</collection><collection>ProQuest Central Korea</collection><collection>Health Research Premium Collection</collection><collection>Health Research Premium Collection (Alumni)</collection><collection>ProQuest Health & Medical Complete (Alumni)</collection><collection>Health & Medical Collection (Alumni Edition)</collection><collection>Publicly Available Content Database</collection><collection>ProQuest One Academic Eastern Edition (DO NOT USE)</collection><collection>ProQuest One Academic</collection><collection>ProQuest One Academic UKI Edition</collection><collection>ProQuest Central China</collection><collection>PubMed Central (Full Participant titles)</collection><collection>DOAJ Directory of Open Access Journals</collection><jtitle>Orthopaedic surgery</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Meng, Chen‐yang</au><au>Xue, Fei</au><au>Zhao, Zhen‐qun</au><au>Hao, Ting</au><au>Guo, Shi‐bing</au><au>Feng, Wei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Influence of MicroRNA‐141 on Inhibition of the Proliferation of Bone Marrow Mesenchymal Stem Cells in Steroid‐Induced Osteonecrosis via SOX11</atitle><jtitle>Orthopaedic surgery</jtitle><stitle>ORTHOP SURG</stitle><addtitle>Orthop Surg</addtitle><date>2020-02</date><risdate>2020</risdate><volume>12</volume><issue>1</issue><spage>277</spage><epage>285</epage><pages>277-285</pages><issn>1757-7853</issn><eissn>1757-7861</eissn><abstract><![CDATA[Objective
To investigate whether miR‐141 and the sex determination region of Y chromosome box 11 (SOX11) play roles in steroid‐induced avascular necrosis of the femoral head (SANFH), and to explore whether miR‐141 could target SOX11 to influence the proliferation of bone marrow mesenchymal stem cells (BMSC).
Methods
Bone marrow mesenchymal stem cells (BMSC) were isolated and cultured from 4‐week‐old Sprague Dawley rats. A flow cytometry assay was performed to identify BMSC. BMSC were divided into two groups: a control group and a dexamethasone (DEX) group. BMSC were transfected by miR‐141 mimic, miR‐141 inhibitor, and SOX11. Real‐time polymerase chain reaction (PCR) assay was performed to investigate the mRNA expression of miR‐141 and SOX11. The results were used to determine the effect of transfection and to verify the expression in each group and the association between miR‐141 and SOX11. Luciferase reporter assay revealed the targeted binding site between miR‐141 and the 3′‐untranslated region of SOX11 mRNA. MTT assays were performed to investigate the proliferation of BMSC in the miR‐141 mimic, miR‐141 inhibitor, and SOX11 groups.
Result
The results of the flow cytometry assay suggested that cells were positive for CD29 and CD90 while negative for CD45. This meant that the isolated and cultured cells were not hematopoietic stem cells. In addition, cell transfection was successful based on the expression of miR‐141 and SOX11. According to the results of real‐time PCR assay, the mRNA expression of miR‐141 in SANFH was upregulated (4.117 ± 0.042 vs 1 ± 0.027, P < 0.001), while SOX11 was downregulated (0.611 ± 0.055 vs 1 ± 0.027, P < 0.001) compared with the control group. Based on the results of the luciferase experiment, MiR‐141 could directly target the expression of SOX11. Inhibition of miR‐141 could upregulate the expression of SOX11 (2.623 ± 0.220 vs 1 ± 0.095, P < 0.001) according to the results of a real‐time PCR assay. MiR‐141 inhibited the proliferation of BMSC (0.618 ± 0.092 vs 1.004 ± 0.082, P < 0.001), while suppression of miR‐141 increased the proliferation of BMSC (0.960 ± 0.095 vs 0.742 ± 0.091, P < 0.001). Furthermore, according to the results of the MTT assay, SOX11 promoted the proliferation of BMSC (1.064 ± 0.093 vs 0.747 ± 0.090, P < 0.001).
Conclusion
MiR‐141 inhibited the proliferation of BMSC in SANFH by targeting SOX11. Inhibition of miR‐141 upregulated the expression of SOX11 and promoted the proliferation of BMSC. MiR‐141 and SOX11 could be new targets for investigating the mechanism of SANFH.]]></abstract><cop>Melbourne</cop><pub>John Wiley & Sons Australia, Ltd</pub><pmid>31916393</pmid><doi>10.1111/os.12603</doi><tpages>9</tpages><orcidid>https://orcid.org/0000-0001-7913-2011</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Adipocytes Bone marrow Bone marrow mesenchymal stem cells Cell culture Cell growth Flow cytometry Gene expression Glucocorticoid Life Sciences & Biomedicine Lipids Metabolism MicroRNAs MicroRNA‐141 Monoclonal antibodies Orthopedics Osteoporosis Polymerase chain reaction Science & Technology Scientific SOX11 Stem cells Steroid induced avascular necrosis of femoral head Y chromosomes |
| title | Influence of MicroRNA‐141 on Inhibition of the Proliferation of Bone Marrow Mesenchymal Stem Cells in Steroid‐Induced Osteonecrosis via SOX11 |
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