An Oligodeoxynucleotide with Promising Modulation Activity for the Proliferation and Activation of Osteoblast
Identifieur interne : 002E42 ( Pmc/Curation ); précédent : 002E41; suivant : 002E43An Oligodeoxynucleotide with Promising Modulation Activity for the Proliferation and Activation of Osteoblast
Auteurs : Zhiyuan Feng ; Yuqin Shen ; Liying Wang ; Lin Cheng ; Jing Wang ; Quanshun Li ; Wei Shi ; Xinhua SunSource :
- International Journal of Molecular Sciences [ 1422-0067 ] ; 2011.
Abstract
The paper explored the regulatory role of oligodeoxynucleotides (ODNs) with specific sequences in the proliferation and activation of osteoblast, using human osteoblast-like cell line MG 63 as the model. Through the administration of ODNs to MG 63 cells at a concentration of 1.0 μg/mL, ODN MT01 with positive effects on proliferation and activation of osteoblast was selected from 11 different ODNs by methyl thiazolyl tetrazolium (MTT) assay and alkaline phosphatase (ALP) activity measurement. To get a deeper insight into the molecular mechanism, effects of ODN MT01 treatment on the expression level of Sp7, runx-2, collagen-I, osteoprotegerin (OPG) and RANK ligand (RANKL) were determined using quantitative real time PCR and Western blotting. Remarkably, the mRNA and protein expression levels of Sp7, runx-2, collagen-I and OPG were improved after ODN MT01 treatment. Meanwhile, the protein expression level of RANKL was dramatically decreased. These results suggested that ODN MT01 had a significant impact in facilitating osteogenic proliferation and activation, and provided a direct evidence for the notion that single strand ODN could regulate the balance of bone formation and resorption, and thus was of great potential in the rebuilding of alveolar bone.
Url:
DOI: 10.3390/ijms12042543
PubMed: 21731457
PubMed Central: 3127133
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">An Oligodeoxynucleotide with Promising Modulation Activity for the Proliferation and Activation of Osteoblast</title><author><name sortKey="Feng, Zhiyuan" sort="Feng, Zhiyuan" uniqKey="Feng Z" first="Zhiyuan" last="Feng">Zhiyuan Feng</name><affiliation><nlm:aff id="af1-ijms-12-02543">Department of Orthodontics, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mails:<email>fzy00@sina.com</email> (Z.F.);<email>33817805@qq.com</email> (J.W.)</nlm:aff></affiliation></author><author><name sortKey="Shen, Yuqin" sort="Shen, Yuqin" uniqKey="Shen Y" first="Yuqin" last="Shen">Yuqin Shen</name><affiliation><nlm:aff id="af2-ijms-12-02543">Department of Periodontal, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mail:<email>shenyuqin326@sina.com</email></nlm:aff></affiliation></author><author><name sortKey="Wang, Liying" sort="Wang, Liying" uniqKey="Wang L" first="Liying" last="Wang">Liying Wang</name><affiliation><nlm:aff id="af3-ijms-12-02543">Department of Molecular Biology, College of Basic Medicine, Jilin University, Changchun 130021, China; E-Mail:<email>wlying@jlu.edu.cn</email></nlm:aff></affiliation></author><author><name sortKey="Cheng, Lin" sort="Cheng, Lin" uniqKey="Cheng L" first="Lin" last="Cheng">Lin Cheng</name><affiliation><nlm:aff id="af4-ijms-12-02543">Department of Oral Cavity, The Second Affiliated Hospital of Shanxi Medical University, 382 Wuyi Road, Taiyuan 030001, China; E-Mail:<email>chenglin6002@sina.com</email></nlm:aff></affiliation></author><author><name sortKey="Wang, Jing" sort="Wang, Jing" uniqKey="Wang J" first="Jing" last="Wang">Jing Wang</name><affiliation><nlm:aff id="af1-ijms-12-02543">Department of Orthodontics, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mails:<email>fzy00@sina.com</email> (Z.F.);<email>33817805@qq.com</email> (J.W.)</nlm:aff></affiliation></author><author><name sortKey="Li, Quanshun" sort="Li, Quanshun" uniqKey="Li Q" first="Quanshun" last="Li">Quanshun Li</name><affiliation><nlm:aff id="af5-ijms-12-02543">Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, 2699 Qianjin Road, Changchun 130021, China; 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E-Mails:<email>fzy00@sina.com</email> (Z.F.);<email>33817805@qq.com</email> (J.W.)</nlm:aff></affiliation></author><author><name sortKey="Shen, Yuqin" sort="Shen, Yuqin" uniqKey="Shen Y" first="Yuqin" last="Shen">Yuqin Shen</name><affiliation><nlm:aff id="af2-ijms-12-02543">Department of Periodontal, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mail:<email>shenyuqin326@sina.com</email></nlm:aff></affiliation></author><author><name sortKey="Wang, Liying" sort="Wang, Liying" uniqKey="Wang L" first="Liying" last="Wang">Liying Wang</name><affiliation><nlm:aff id="af3-ijms-12-02543">Department of Molecular Biology, College of Basic Medicine, Jilin University, Changchun 130021, China; E-Mail:<email>wlying@jlu.edu.cn</email></nlm:aff></affiliation></author><author><name sortKey="Cheng, Lin" sort="Cheng, Lin" uniqKey="Cheng L" first="Lin" last="Cheng">Lin Cheng</name><affiliation><nlm:aff id="af4-ijms-12-02543">Department of Oral Cavity, The Second Affiliated Hospital of Shanxi Medical University, 382 Wuyi Road, Taiyuan 030001, China; E-Mail:<email>chenglin6002@sina.com</email></nlm:aff></affiliation></author><author><name sortKey="Wang, Jing" sort="Wang, Jing" uniqKey="Wang J" first="Jing" last="Wang">Jing Wang</name><affiliation><nlm:aff id="af1-ijms-12-02543">Department of Orthodontics, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mails:<email>fzy00@sina.com</email> (Z.F.);<email>33817805@qq.com</email> (J.W.)</nlm:aff></affiliation></author><author><name sortKey="Li, Quanshun" sort="Li, Quanshun" uniqKey="Li Q" first="Quanshun" last="Li">Quanshun Li</name><affiliation><nlm:aff id="af5-ijms-12-02543">Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, 2699 Qianjin Road, Changchun 130021, China; E-Mail:<email>quanshun@jlu.edu.cn</email></nlm:aff></affiliation></author><author><name sortKey="Shi, Wei" sort="Shi, Wei" uniqKey="Shi W" first="Wei" last="Shi">Wei Shi</name><affiliation><nlm:aff id="af5-ijms-12-02543">Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, 2699 Qianjin Road, Changchun 130021, China; E-Mail:<email>quanshun@jlu.edu.cn</email></nlm:aff></affiliation></author><author><name sortKey="Sun, Xinhua" sort="Sun, Xinhua" uniqKey="Sun X" first="Xinhua" last="Sun">Xinhua Sun</name><affiliation><nlm:aff id="af1-ijms-12-02543">Department of Orthodontics, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mails:<email>fzy00@sina.com</email> (Z.F.);<email>33817805@qq.com</email> (J.W.)</nlm:aff></affiliation></author></analytic><series><title level="j">International Journal of Molecular Sciences</title><idno type="eISSN">1422-0067</idno><imprint><date when="2011">2011</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>The paper explored the regulatory role of oligodeoxynucleotides (ODNs) with specific sequences in the proliferation and activation of osteoblast, using human osteoblast-like cell line MG 63 as the model. Through the administration of ODNs to MG 63 cells at a concentration of 1.0 μg/mL, ODN MT01 with positive effects on proliferation and activation of osteoblast was selected from 11 different ODNs by methyl thiazolyl tetrazolium (MTT) assay and alkaline phosphatase (ALP) activity measurement. To get a deeper insight into the molecular mechanism, effects of ODN MT01 treatment on the expression level of Sp7, runx-2, collagen-I, osteoprotegerin (OPG) and RANK ligand (RANKL) were determined using quantitative real time PCR and Western blotting. Remarkably, the mRNA and protein expression levels of Sp7, runx-2, collagen-I and OPG were improved after ODN MT01 treatment. Meanwhile, the protein expression level of RANKL was dramatically decreased. These results suggested that ODN MT01 had a significant impact in facilitating osteogenic proliferation and activation, and provided a direct evidence for the notion that single strand ODN could regulate the balance of bone formation and resorption, and thus was of great potential in the rebuilding of alveolar bone.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Verna, C" uniqKey="Verna C">C Verna</name></author><author><name sortKey="Zaffe, D" uniqKey="Zaffe D">D Zaffe</name></author><author><name sortKey="Siciliani, G" uniqKey="Siciliani G">G Siciliani</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Graves, Dt" uniqKey="Graves D">DT Graves</name></author><author><name sortKey="Kayal, Ra" uniqKey="Kayal R">RA Kayal</name></author><author><name sortKey="Oates, T" uniqKey="Oates T">T Oates</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Ducy, P" uniqKey="Ducy 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<front><journal-meta><journal-id journal-id-type="nlm-ta">Int J Mol Sci</journal-id><journal-id journal-id-type="publisher-id">ijms</journal-id><journal-title-group><journal-title>International Journal of Molecular Sciences</journal-title></journal-title-group><issn pub-type="epub">1422-0067</issn><publisher><publisher-name>Molecular Diversity Preservation International (MDPI)</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">21731457</article-id><article-id pub-id-type="pmc">3127133</article-id><article-id pub-id-type="doi">10.3390/ijms12042543</article-id><article-id pub-id-type="publisher-id">ijms-12-02543</article-id><article-categories><subj-group subj-group-type="heading"><subject>Article</subject></subj-group></article-categories><title-group><article-title>An Oligodeoxynucleotide with Promising Modulation Activity for the Proliferation and Activation of Osteoblast</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Feng</surname><given-names>Zhiyuan</given-names></name><xref ref-type="aff" rid="af1-ijms-12-02543"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Shen</surname><given-names>Yuqin</given-names></name><xref ref-type="aff" rid="af2-ijms-12-02543"><sup>2</sup></xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Liying</given-names></name><xref ref-type="aff" rid="af3-ijms-12-02543"><sup>3</sup></xref></contrib><contrib contrib-type="author"><name><surname>Cheng</surname><given-names>Lin</given-names></name><xref ref-type="aff" rid="af4-ijms-12-02543"><sup>4</sup></xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Jing</given-names></name><xref ref-type="aff" rid="af1-ijms-12-02543"><sup>1</sup></xref></contrib><contrib contrib-type="author"><name><surname>Li</surname><given-names>Quanshun</given-names></name><xref ref-type="aff" rid="af5-ijms-12-02543"><sup>5</sup></xref></contrib><contrib contrib-type="author"><name><surname>Shi</surname><given-names>Wei</given-names></name><xref ref-type="aff" rid="af5-ijms-12-02543"><sup>5</sup></xref><xref ref-type="corresp" rid="c1-ijms-12-02543"><sup>*</sup></xref></contrib><contrib contrib-type="author"><name><surname>Sun</surname><given-names>Xinhua</given-names></name><xref ref-type="aff" rid="af1-ijms-12-02543"><sup>1</sup></xref><xref ref-type="corresp" rid="c1-ijms-12-02543"><sup>*</sup></xref></contrib></contrib-group><aff id="af1-ijms-12-02543"><label>1</label>Department of Orthodontics, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mails:<email>fzy00@sina.com</email> (Z.F.);<email>33817805@qq.com</email> (J.W.)</aff><aff id="af2-ijms-12-02543"><label>2</label>Department of Periodontal, School of Stomatology, Jilin University, 1500 Qinghua Road, Changchun 130021, China; E-Mail:<email>shenyuqin326@sina.com</email></aff><aff id="af3-ijms-12-02543"><label>3</label>Department of Molecular Biology, College of Basic Medicine, Jilin University, Changchun 130021, China; E-Mail:<email>wlying@jlu.edu.cn</email></aff><aff id="af4-ijms-12-02543"><label>4</label>Department of Oral Cavity, The Second Affiliated Hospital of Shanxi Medical University, 382 Wuyi Road, Taiyuan 030001, China; E-Mail:<email>chenglin6002@sina.com</email></aff><aff id="af5-ijms-12-02543"><label>5</label>Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, Jilin University, 2699 Qianjin Road, Changchun 130021, China; E-Mail:<email>quanshun@jlu.edu.cn</email></aff><author-notes><corresp id="c1-ijms-12-02543"><label>*</label>Authors to whom correspondence should be addressed; E-Mails: <email>shiw@jlu.edu.cn</email> (W.S.); <email>xinhuasun8@163.com</email> (X.S.); Tel.: +86-431-88796023; Fax: +86-431-88975348.</corresp></author-notes><pub-date pub-type="epub"><day>15</day><month>4</month><year>2011</year></pub-date><pub-date pub-type="collection"><year>2011</year></pub-date><volume>12</volume><issue>4</issue><fpage>2543</fpage><lpage>2555</lpage><history><date date-type="received"><day>11</day><month>3</month><year>2011</year></date><date date-type="rev-recd"><day>6</day><month>4</month><year>2011</year></date><date date-type="accepted"><day>7</day><month>4</month><year>2011</year></date></history><permissions><copyright-statement>© 2011 by the authors; licensee MDPI, Basel, Switzerland.</copyright-statement><copyright-year>2011</copyright-year><license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/3.0"><license-p><pmc-comment>CREATIVE COMMONS</pmc-comment>This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (<ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/3.0/">http://creativecommons.org/licenses/by/3.0/</ext-link>).</license-p></license></permissions><abstract><p>The paper explored the regulatory role of oligodeoxynucleotides (ODNs) with specific sequences in the proliferation and activation of osteoblast, using human osteoblast-like cell line MG 63 as the model. Through the administration of ODNs to MG 63 cells at a concentration of 1.0 μg/mL, ODN MT01 with positive effects on proliferation and activation of osteoblast was selected from 11 different ODNs by methyl thiazolyl tetrazolium (MTT) assay and alkaline phosphatase (ALP) activity measurement. To get a deeper insight into the molecular mechanism, effects of ODN MT01 treatment on the expression level of Sp7, runx-2, collagen-I, osteoprotegerin (OPG) and RANK ligand (RANKL) were determined using quantitative real time PCR and Western blotting. Remarkably, the mRNA and protein expression levels of Sp7, runx-2, collagen-I and OPG were improved after ODN MT01 treatment. Meanwhile, the protein expression level of RANKL was dramatically decreased. These results suggested that ODN MT01 had a significant impact in facilitating osteogenic proliferation and activation, and provided a direct evidence for the notion that single strand ODN could regulate the balance of bone formation and resorption, and thus was of great potential in the rebuilding of alveolar bone.</p></abstract><kwd-group><kwd>oligodeoxynucleotide</kwd><kwd>osteoblast</kwd><kwd>proliferation</kwd><kwd>activation</kwd><kwd>bone remodeling</kwd></kwd-group></article-meta></front><floats-group><fig id="f1-ijms-12-02543" position="float"><label>Figure 1.</label><caption><p>Effects of 11 different ODNs on the cell viability and proliferation of MG 63 osteoblast-like cells after 72 h treatment. *Statistically significant difference (<italic>P</italic> < 0.05) between experimental and control groups.</p></caption><graphic xlink:href="ijms-12-02543f1"></graphic></fig><fig id="f2-ijms-12-02543" position="float"><label>Figure 2.</label><caption><p>Effect of ODNs treatment on ALP activity of MG 63 cells, for different time intervals. *Statistically significant difference (<italic>P</italic> < 0.05) between experimental and control groups at the given time point.</p></caption><graphic xlink:href="ijms-12-02543f2"></graphic></fig><fig id="f3-ijms-12-02543" position="float"><label>Figure 3.</label><caption><p>(<bold>A</bold>) MG 63 cells cultured in DMEM for 72 h; (<bold>B</bold>) MG 63 cells cultured in ODN MT01 (1.0 μg/mL)-DMEM for 72 h.</p></caption><graphic xlink:href="ijms-12-02543f3"></graphic></fig><fig id="f4-ijms-12-02543" position="float"><label>Figure 4.</label><caption><p>Real time PCR analysis of MG 63 cells of experimental group (ODN MT01 treatment) and control group (PBS treatment) at predetermined time. (<bold>A</bold>) Relative amounts of mRNA for Sp7 were quantified and GAPDH was used as internal control; (<bold>B</bold>) Relative amounts of mRNA for runx-2 were quantified and GAPDH was used as internal control; (<bold>C</bold>) Relative amounts of mRNA for collagen-I were quantified and GAPDH was used as internal control. *Statistically significant difference (<italic>P</italic> < 0.05) between experimental and control groups (<italic>n</italic> = 3).</p></caption><graphic xlink:href="ijms-12-02543f4a"></graphic><graphic xlink:href="ijms-12-02543f4b"></graphic></fig><fig id="f5-ijms-12-02543" position="float"><label>Figure 5.</label><caption><p>Western blotting analysis of expression level of Sp7, runx-2, collagen-I, OPG and RANKL in ODN MT01 treatment group and control group at predetermined time.</p></caption><graphic xlink:href="ijms-12-02543f5"></graphic></fig><table-wrap id="t1-ijms-12-02543" position="float"><label>Table 1.</label><caption><p>ODNs (11 different sequences) were used for co-culture with MG 63 cells to screen the ones that could improve the proliferation of cells.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="center" valign="bottom" rowspan="1" colspan="1"><bold>Number</bold></th><th align="center" valign="bottom" rowspan="1" colspan="1"><bold>Name</bold></th><th align="center" valign="bottom" rowspan="1" colspan="1"><bold>Sequence</bold></th></tr></thead><tbody><tr><td align="center" valign="top" rowspan="1" colspan="1">1</td><td align="center" valign="top" rowspan="1" colspan="1">SAT05f</td><td align="center" valign="top" rowspan="1" colspan="1">5′-CCTCCTCCTCCTCCTCCTCCTCCT-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">2</td><td align="center" valign="top" rowspan="1" colspan="1">MS19</td><td align="center" valign="top" rowspan="1" colspan="1">5′-AAAGAAAGAAAGAAAGAAAGAAAG-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">3</td><td align="center" valign="top" rowspan="1" colspan="1">BW001</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGTCGGGTGCGACGTCGCAGGGGGG-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">4</td><td align="center" valign="top" rowspan="1" colspan="1">FC001</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGGGGACGATCGTCGGGGAC-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">5</td><td align="center" valign="top" rowspan="1" colspan="1">FC002</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGTCGACGTCGTCGTTCTC-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">6</td><td align="center" valign="top" rowspan="1" colspan="1">BW006</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGACGTTCGTCGTTCGTCGTTC-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">7</td><td align="center" valign="top" rowspan="1" colspan="1">YW002</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGCGAACGTTCGCCGCGTTCGAACGCGG-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">8</td><td align="center" valign="top" rowspan="1" colspan="1">FC004</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGCGTTCGATCGCGATCGACGGTA-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">9</td><td align="center" valign="top" rowspan="1" colspan="1">MT01</td><td align="center" valign="top" rowspan="1" colspan="1">5′-ACCCCCTCTACCCCCTCTACCCCCTCT-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">10</td><td align="center" valign="top" rowspan="1" colspan="1">YW001</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCGCGACGTTCGCCCGACGTTCGGTA-3′</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">11</td><td align="center" valign="top" rowspan="1" colspan="1">FC003</td><td align="center" valign="top" rowspan="1" colspan="1">5′-TCTCTCTCTCTCTCTCTCTCTCTC-3′</td></tr></tbody></table></table-wrap><table-wrap id="t2-ijms-12-02543" position="float"><label>Table 2.</label><caption><p>The primers used for qRT-PCR of various genes.</p></caption><table frame="hsides" rules="groups"><thead><tr><th align="center" valign="middle" rowspan="1" colspan="1"><bold>Gene Name</bold></th><th align="center" valign="middle" rowspan="1" colspan="1"><bold>Oligonucleotide UP (5′-3′)</bold></th><th align="center" valign="middle" rowspan="1" colspan="1"><bold>Oligonucleotide DW (5′-3′)</bold></th></tr></thead><tbody><tr><td align="center" valign="top" rowspan="1" colspan="1">GAPDH</td><td align="center" valign="top" rowspan="1" colspan="1">ATG GGG AAG GTG AAG GTC</td><td align="center" valign="top" rowspan="1" colspan="1">TAA AAG CAG CCC TGG TGA CC</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">SP7</td><td align="center" valign="top" rowspan="1" colspan="1">CAC AGC TCT TCT GAC TGT CTG CTG</td><td align="center" valign="top" rowspan="1" colspan="1">GTG AAA TGC CTG CAT GGA T</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">RUNX-2</td><td align="center" valign="top" rowspan="1" colspan="1">GAG ATC ATC GCC GAC CAC</td><td align="center" valign="top" rowspan="1" colspan="1">TAC CTC TCC GAG GGC TAC C</td></tr><tr><td align="center" valign="top" rowspan="1" colspan="1">COLLAGEN-I</td><td align="center" valign="top" rowspan="1" colspan="1">AGG GCC AAG ACG AAG ACA TC</td><td align="center" valign="top" rowspan="1" colspan="1">AGA TCA CGT CAT CGC ACA ACA</td></tr></tbody></table></table-wrap></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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