Saikosaponin-d Increases the Radiosensitivity of Hepatoma Cells by Adjusting Cell Autophagy
Identifieur interne : 000647 ( Pmc/Curation ); précédent : 000646; suivant : 000648Saikosaponin-d Increases the Radiosensitivity of Hepatoma Cells by Adjusting Cell Autophagy
Auteurs : Yin-Di Tian [République populaire de Chine] ; Shuai Lin [République populaire de Chine] ; Peng-Tao Yang [République populaire de Chine] ; Ming-Hua Bai [République populaire de Chine] ; Ying-Ying Jin [République populaire de Chine] ; Wei-Li Min [République populaire de Chine] ; Hong-Bing Ma [République populaire de Chine] ; Bao-Feng Wang [République populaire de Chine]Source :
- Journal of Cancer [ 1837-9664 ] ; 2019.
Abstract
Radiotherapy for liver cancer can affect the level of autophagy in cells, and effective autophagy regulation can increase the radiosensitivity of liver cancer cells.Saikosaponin-d (SSd) is an effective active ingredient extracted from traditional Chinese medicine Bupleurum. We have confirmed previously in vitro and in vitro experiments that SSd can significantly induce apoptosis of liver cancer cells, increase the radiosensitivity of liver cancer cells.This study explored the role of autophagy in SSd-mediated radiosensitivity of liver cancer cells. MTT and clone formation experiments showed that radiation can inhibit the proliferation of hepatoma cells and reduce the colony formation of hepatoma cells. After the addition of SSd, the inhibitory effect of radiation on the proliferation and clonal formation of hepatoma cells was further enhanced. However, the addition of the autophagy inhibitor chloroquine or mTOR agonist can partially reverse the inhibitory effect of the combined treatment of SSd with radiation on the proliferation of hepatoma cells. Similarly, transmission electron microscopy and laser confocal microscopy showed that after the addition of SSd, the number of radiation-induced autophagosomes increased significantly in hepatoma cells and the intervention of mTOR agonist can reduce the formation of autophagosomes in hepatoma cells.In addition,Western blot analysis presented that radiation significantly increased LC3-II levels. Especially when SSd is added, LC3-II levels is further increased. Our data indicate that SSd can inhibit the growth of liver cancer cells and enhance cell radiosensitivity by inducing autophagy formation.
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DOI: 10.7150/jca.30286
PubMed: 31598167
PubMed Central: 6775525
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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Saikosaponin-d Increases the Radiosensitivity of Hepatoma Cells by Adjusting Cell Autophagy</title><author><name sortKey="Tian, Yin Di" sort="Tian, Yin Di" uniqKey="Tian Y" first="Yin-Di" last="Tian">Yin-Di Tian</name><affiliation wicri:level="1"><nlm:aff id="A1">Department of Infectious Diseases, the Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Infectious Diseases, the Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Lin, Shuai" sort="Lin, Shuai" uniqKey="Lin S" first="Shuai" last="Lin">Shuai Lin</name><affiliation wicri:level="1"><nlm:aff id="A2">Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Yang, Peng Tao" sort="Yang, Peng Tao" uniqKey="Yang P" first="Peng-Tao" last="Yang">Peng-Tao Yang</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Bai, Ming Hua" sort="Bai, Ming Hua" uniqKey="Bai M" first="Ming-Hua" last="Bai">Ming-Hua Bai</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Jin, Ying Ying" sort="Jin, Ying Ying" uniqKey="Jin Y" first="Ying-Ying" last="Jin">Ying-Ying Jin</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Min, Wei Li" sort="Min, Wei Li" uniqKey="Min W" first="Wei-Li" last="Min">Wei-Li Min</name><affiliation wicri:level="1"><nlm:aff id="A2">Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Ma, Hong Bing" sort="Ma, Hong Bing" uniqKey="Ma H" first="Hong-Bing" last="Ma">Hong-Bing Ma</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Bao Feng" sort="Wang, Bao Feng" uniqKey="Wang B" first="Bao-Feng" last="Wang">Bao-Feng Wang</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">31598167</idno><idno type="pmc">6775525</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6775525</idno><idno type="RBID">PMC:6775525</idno><idno type="doi">10.7150/jca.30286</idno><date when="2019">2019</date><idno type="wicri:Area/Pmc/Corpus">000647</idno><idno type="wicri:explorRef" 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Shuai" uniqKey="Lin S" first="Shuai" last="Lin">Shuai Lin</name><affiliation wicri:level="1"><nlm:aff id="A2">Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Yang, Peng Tao" sort="Yang, Peng Tao" uniqKey="Yang P" first="Peng-Tao" last="Yang">Peng-Tao Yang</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Bai, Ming Hua" sort="Bai, Ming Hua" uniqKey="Bai M" first="Ming-Hua" last="Bai">Ming-Hua Bai</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Jin, Ying Ying" sort="Jin, Ying Ying" uniqKey="Jin Y" first="Ying-Ying" last="Jin">Ying-Ying Jin</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Min, Wei Li" sort="Min, Wei Li" uniqKey="Min W" first="Wei-Li" last="Min">Wei-Li Min</name><affiliation wicri:level="1"><nlm:aff id="A2">Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Ma, Hong Bing" sort="Ma, Hong Bing" uniqKey="Ma H" first="Hong-Bing" last="Ma">Hong-Bing Ma</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author><author><name sortKey="Wang, Bao Feng" sort="Wang, Bao Feng" uniqKey="Wang B" first="Bao-Feng" last="Wang">Bao-Feng Wang</name><affiliation wicri:level="1"><nlm:aff id="A3">Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</nlm:aff><country xml:lang="fr">République populaire de Chine</country><wicri:regionArea>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004</wicri:regionArea></affiliation></author></analytic><series><title level="j">Journal of Cancer</title><idno type="eISSN">1837-9664</idno><imprint><date when="2019">2019</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>Radiotherapy for liver cancer can affect the level of autophagy in cells, and effective autophagy regulation can increase the radiosensitivity of liver cancer cells.Saikosaponin-d (SSd) is an effective active ingredient extracted from traditional Chinese medicine Bupleurum. We have confirmed previously in vitro and in vitro experiments that SSd can significantly induce apoptosis of liver cancer cells, increase the radiosensitivity of liver cancer cells.This study explored the role of autophagy in SSd-mediated radiosensitivity of liver cancer cells. MTT and clone formation experiments showed that radiation can inhibit the proliferation of hepatoma cells and reduce the colony formation of hepatoma cells. After the addition of SSd, the inhibitory effect of radiation on the proliferation and clonal formation of hepatoma cells was further enhanced. However, the addition of the autophagy inhibitor chloroquine or mTOR agonist can partially reverse the inhibitory effect of the combined treatment of SSd with radiation on the proliferation of hepatoma cells. Similarly, transmission electron microscopy and laser confocal microscopy showed that after the addition of SSd, the number of radiation-induced autophagosomes increased significantly in hepatoma cells and the intervention of mTOR agonist can reduce the formation of autophagosomes in hepatoma cells.In addition,Western blot analysis presented that radiation significantly increased LC3-II levels. Especially when SSd is added, LC3-II levels is further increased. Our data indicate that SSd can inhibit the growth of liver cancer cells and enhance cell radiosensitivity by inducing autophagy formation.</p></div></front><back><div1 type="bibliography"><listBibl><biblStruct><analytic><author><name sortKey="Kim, Dy" uniqKey="Kim D">DY Kim</name></author><author><name sortKey="Han, Kh" uniqKey="Han K">KH Han</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Cong, Wm" uniqKey="Cong W">WM Cong</name></author><author><name sortKey="Bu, H" uniqKey="Bu H">H Bu</name></author><author><name sortKey="Chen, J" uniqKey="Chen J">J Chen</name></author><author><name sortKey="Guideline, Committee" uniqKey="Guideline C">Committee Guideline</name></author></analytic></biblStruct><biblStruct><analytic><author><name sortKey="Jemal, A" uniqKey="Jemal A">A Jemal</name></author><author><name sortKey="Bray, F" uniqKey="Bray F">F Bray</name></author><author><name sortKey="Center, Mm" uniqKey="Center M">MM 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<front><journal-meta><journal-id journal-id-type="nlm-ta">J Cancer</journal-id><journal-id journal-id-type="iso-abbrev">J Cancer</journal-id><journal-id journal-id-type="publisher-id">jca</journal-id><journal-title-group><journal-title>Journal of Cancer</journal-title></journal-title-group><issn pub-type="epub">1837-9664</issn><publisher><publisher-name>Ivyspring International Publisher</publisher-name><publisher-loc>Sydney</publisher-loc></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">31598167</article-id><article-id pub-id-type="pmc">6775525</article-id><article-id pub-id-type="doi">10.7150/jca.30286</article-id><article-id pub-id-type="publisher-id">jcav10p4947</article-id><article-categories><subj-group subj-group-type="heading"><subject>Research Paper</subject></subj-group></article-categories><title-group><article-title>Saikosaponin-d Increases the Radiosensitivity of Hepatoma Cells by Adjusting Cell Autophagy</article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Tian</surname><given-names>Yin-Di</given-names></name><xref ref-type="aff" rid="A1">1</xref><xref ref-type="author-notes" rid="FNA_number">#</xref></contrib><contrib contrib-type="author"><name><surname>Lin</surname><given-names>Shuai</given-names></name><xref ref-type="aff" rid="A2">2</xref><xref ref-type="author-notes" rid="FNA_number">#</xref></contrib><contrib contrib-type="author"><name><surname>Yang</surname><given-names>Peng-Tao</given-names></name><xref ref-type="aff" rid="A3">3</xref><xref ref-type="author-notes" rid="FNA_number">#</xref></contrib><contrib contrib-type="author"><name><surname>Bai</surname><given-names>Ming-Hua</given-names></name><xref ref-type="aff" rid="A3">3</xref></contrib><contrib contrib-type="author"><name><surname>Jin</surname><given-names>Ying-Ying</given-names></name><xref ref-type="aff" rid="A3">3</xref></contrib><contrib contrib-type="author"><name><surname>Min</surname><given-names>Wei-Li</given-names></name><xref ref-type="aff" rid="A2">2</xref></contrib><contrib contrib-type="author"><name><surname>Ma</surname><given-names>Hong-Bing</given-names></name><xref ref-type="aff" rid="A3">3</xref><xref ref-type="corresp" rid="FNA_envelop">✉</xref></contrib><contrib contrib-type="author"><name><surname>Wang</surname><given-names>Bao-Feng</given-names></name><xref ref-type="aff" rid="A3">3</xref><xref ref-type="corresp" rid="FNA_envelop">✉</xref></contrib></contrib-group><aff id="A1"><label>1</label>Department of Infectious Diseases, the Second Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an 710004, China</aff><aff id="A2"><label>2</label>Department of Surgical Oncology, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</aff><aff id="A3"><label>3</label>Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China</aff><author-notes><corresp id="FNA_envelop">✉ Corresponding author: <bold>Address:</bold> Department of Radiation Therapy, Second Affiliated Hospital of Medical School, Xi'an Jiaotong University, Xi'an 710004, China. Tel: +86 29 87679789; E-mail:<email>mhbxidn@126.com</email>; <email>wangbf1680@126.com</email></corresp><fn fn-type="equal" id="FNA_number"><p><sup>#</sup>These authors contribute equally to this work.</p></fn><fn fn-type="COI-statement"><p>Competing Interests: The authors have declared that no competing interest exists.</p></fn></author-notes><pub-date pub-type="collection"><year>2019</year></pub-date><pub-date pub-type="epub"><day>27</day><month>8</month><year>2019</year></pub-date><volume>10</volume><issue>20</issue><fpage>4947</fpage><lpage>4953</lpage><history><date date-type="received"><day>29</day><month>9</month><year>2018</year></date><date date-type="accepted"><day>23</day><month>6</month><year>2019</year></date></history><permissions><copyright-statement>© The author(s)</copyright-statement><copyright-year>2019</copyright-year><license license-type="open-access"><license-p>This is an open access article distributed under the terms of the Creative Commons Attribution License (<ext-link ext-link-type="uri" xlink:href="https://creativecommons.org/licenses/by/4.0/">https://creativecommons.org/licenses/by/4.0/</ext-link>). See <ext-link ext-link-type="uri" xlink:href="http://ivyspring.com/terms">http://ivyspring.com/terms</ext-link> for full terms and conditions.</license-p></license></permissions><abstract><p>Radiotherapy for liver cancer can affect the level of autophagy in cells, and effective autophagy regulation can increase the radiosensitivity of liver cancer cells.Saikosaponin-d (SSd) is an effective active ingredient extracted from traditional Chinese medicine Bupleurum. We have confirmed previously in vitro and in vitro experiments that SSd can significantly induce apoptosis of liver cancer cells, increase the radiosensitivity of liver cancer cells.This study explored the role of autophagy in SSd-mediated radiosensitivity of liver cancer cells. MTT and clone formation experiments showed that radiation can inhibit the proliferation of hepatoma cells and reduce the colony formation of hepatoma cells. After the addition of SSd, the inhibitory effect of radiation on the proliferation and clonal formation of hepatoma cells was further enhanced. However, the addition of the autophagy inhibitor chloroquine or mTOR agonist can partially reverse the inhibitory effect of the combined treatment of SSd with radiation on the proliferation of hepatoma cells. Similarly, transmission electron microscopy and laser confocal microscopy showed that after the addition of SSd, the number of radiation-induced autophagosomes increased significantly in hepatoma cells and the intervention of mTOR agonist can reduce the formation of autophagosomes in hepatoma cells.In addition,Western blot analysis presented that radiation significantly increased LC3-II levels. Especially when SSd is added, LC3-II levels is further increased. Our data indicate that SSd can inhibit the growth of liver cancer cells and enhance cell radiosensitivity by inducing autophagy formation.</p></abstract><kwd-group><kwd>Saikosaponin-d</kwd><kwd>Radiation</kwd><kwd>Hepatocellular carcinoma</kwd><kwd>Autophagy.</kwd></kwd-group></article-meta></front><floats-group><fig id="F1" position="float"><label>Figure 1</label><caption><p><bold> Changes in cell viability after different interventions.</bold> A. MTT demonstrate that the effect of different interventions on the viability of SMMC-7721 cells; B. The effect of different interventions on the viability of MHCC97L cells; C. The effect of different interventions on the viability of normal hepatocytes. (*<italic>P</italic>< 0.05,**<italic>P</italic>< 0.01).</p></caption><graphic xlink:href="jcav10p4947g001"></graphic></fig><fig id="F2" position="float"><label>Figure 2</label><caption><p><bold> Changes in clonal formation rate after different interventions.</bold> A more pronounced decrease of colony formation rate was observed in the combined treatment group than that of in radiation groups. Meanwhile, colony formation rate of combined treatment group was increased after the intervention of CQ or MHY1485. A. Colony formation rate changes of SMMC-7721 cells; B. Colony formation rate changes of MHCC97L cells (*<italic>P</italic>< 0.05, **<italic>P</italic>< 0.01).</p></caption><graphic xlink:href="jcav10p4947g002"></graphic></fig><fig id="F3" position="float"><label>Figure 3</label><caption><p><bold> Formation of autophagy in Hepatoma Cells was observed under transmission electron microscope.</bold> Autophagosomes of SMMC-7721 cells in the combined treatment group increased in large quantities, as indicated by the red arrow. However, almost no autophagosomes appeared in MHY1485 groups (×16500).</p></caption><graphic xlink:href="jcav10p4947g003"></graphic></fig><fig id="F4" position="float"><label>Figure 4</label><caption><p><bold> LC3-II expressions were detected by western blot.</bold> A. Western blot analysis of the LC3-II levels in SMMC-7721 cells after treated with different interventions; B. Relative LC3-II expression in SMMC-7721 cells; C. Western blot analysis of the LC3-II levels in SMMC-7721 cells after addition of CQ; D. Relative LC3-II expression in SMMC-7721 cells after addition of CQ. (*<italic>P</italic>< 0.05, **<italic>P</italic>< 0.01).</p></caption><graphic xlink:href="jcav10p4947g004"></graphic></fig><fig id="F5" position="float"><label>Figure 5</label><caption><p><bold> Changes of the number of autophagosomes in SMMC-7721 cells after different interventions.</bold> The number of autophagosomes was 2.33±0.33 in the control group. After the addition of SSd, the number of autophagosomes in the combined group is higher than that of radiation alone group. Representative photographs obtained from each group (A) and statistical analysis from all cells in each group (B). (**<italic>P</italic>< 0.01).</p></caption><graphic xlink:href="jcav10p4947g005"></graphic></fig></floats-group></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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