Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway
BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA topoisomerase.However no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma.This study aimed to investigate the effects of harmine on proliferation and apoptosis and the un...
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| Veröffentlicht in: | Hepatobiliary & pancreatic diseases international 2011-12, Vol.10 (6), p.599-604 |
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| creator | Cao, Ming-Rong Li, Qiang Liu, Zhi-Long Liu, Hui-Hui Wang, Wei Liao, Xiao-Li Pan, Yun-Long Jiang, Jian-Wei |
| description | BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA topoisomerase.However no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma.This study aimed to investigate the effects of harmine on proliferation and apoptosis and the underlying mechanisms in the human hepatocellular carcinoma cell line HepG2.METHODS:The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation test.The morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell cycle.Western blotting was used to assess expression of the apoptosis-regulated genes Bcl-2,Bax,Bcl-xl,Mcl-1,caspase-3,and caspase-9 Mitochondrial transmembrane potential (Ψ m) was determined using JC-1.RESULTS:Harmine inhibited the proliferation of HepG2 cells in a dose-dependent manner.Hoechst 33258 staining revealed nuclear fragmentation and chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with harmine.The percentage of the sub/G1 fraction was increased in a concentration-dependent manner,indicating apoptotic cell death.PI staining showed that harmine changed the cell cycle distribution,by decreasing the proportion of cells inG0/G1 and increasing the proportion in S and G2/M.Harmine induced apoptosis in a concentration-dependent manner,with rates of 20.0%,32.7% and 64.9%,respectively.JC-1 revealed a decrease in Ψ m.Apoptosis of HepG2 cells was associated with caspase-3 and caspase-9 activation,down-regulation of Bcl-2,Mcl-1,and Bcl-xl,and no change in Bax.CONCLUSIONS:Harmine had an anti-proliferative effect in HepG2 cells by inducing apoptosis.Mitochondrial signal pathways were involved in the apoptosis.The cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma. |
| doi_str_mv | 10.1016/S1499-3872(11)60102-1 |
| format | Article |
| fullrecord | <record><control><sourceid>proquest_cross</sourceid><recordid>TN_cdi_proquest_miscellaneous_909754035</recordid><sourceformat>XML</sourceformat><sourcesystem>PC</sourcesystem><cqvip_id>1003032987</cqvip_id><els_id>1_s2_0_S1499387211601021</els_id><sourcerecordid>909754035</sourcerecordid><originalsourceid>FETCH-LOGICAL-c499t-7908e150af6ccffe345784dedbfa19f7ea3fcfac9c08adb2a0c4198bee9526e53</originalsourceid><addsrcrecordid>eNqFkU9v1DAQxX0AtaXlI4AicQAOAY-df75QoQq6SCv1AJytWWe8dUni1E6K9tvj7C4V4tLTSKM3b-b9hrFXwD8Ah-rjdyiUymVTi3cA7ysOXOTwjJ09tk_ZixjvOBdNU1Yn7FQIKKpKyDO2XmHo3UCZG9rZUMxw9OPko4upk61ovBaZoa6L2YPDrHeTN7d-aIPDLotuO2Dnhm024nT7G3cX7LnFLtLLYz1nP79--XG1ytc319-uPq9zkw6a8lrxhqDkaCtjrCVZlHVTtNRuLIKyNaG0xqJRhjfYbgRyU4BqNkSqFBWV8py9PfiOwd_PFCfdu7hciQP5OWrFVV0WXC7K8qA0wccYyOoxuB7DTgPXCzu9Z6cXSBpA79lpSHOvjxvmTU_t49RfcElweRBQyvngKOhoHA2GWhfITLr17skVn_5zMImlM9j9oh3FOz-HRDdq0FFofjBZPAD2DovBm2O29JHtffrDP-G45FKoppZ_AO9KohM</addsrcrecordid><sourcetype>Aggregation Database</sourcetype><iscdi>true</iscdi><recordtype>article</recordtype><pqid>909754035</pqid></control><display><type>article</type><title>Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway</title><source>MEDLINE</source><source>Elsevier ScienceDirect Journals</source><source>Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals</source><creator>Cao, Ming-Rong ; Li, Qiang ; Liu, Zhi-Long ; Liu, Hui-Hui ; Wang, Wei ; Liao, Xiao-Li ; Pan, Yun-Long ; Jiang, Jian-Wei</creator><creatorcontrib>Cao, Ming-Rong ; Li, Qiang ; Liu, Zhi-Long ; Liu, Hui-Hui ; Wang, Wei ; Liao, Xiao-Li ; Pan, Yun-Long ; Jiang, Jian-Wei</creatorcontrib><description>BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA topoisomerase.However no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma.This study aimed to investigate the effects of harmine on proliferation and apoptosis and the underlying mechanisms in the human hepatocellular carcinoma cell line HepG2.METHODS:The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation test.The morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell cycle.Western blotting was used to assess expression of the apoptosis-regulated genes Bcl-2,Bax,Bcl-xl,Mcl-1,caspase-3,and caspase-9 Mitochondrial transmembrane potential (Ψ m) was determined using JC-1.RESULTS:Harmine inhibited the proliferation of HepG2 cells in a dose-dependent manner.Hoechst 33258 staining revealed nuclear fragmentation and chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with harmine.The percentage of the sub/G1 fraction was increased in a concentration-dependent manner,indicating apoptotic cell death.PI staining showed that harmine changed the cell cycle distribution,by decreasing the proportion of cells inG0/G1 and increasing the proportion in S and G2/M.Harmine induced apoptosis in a concentration-dependent manner,with rates of 20.0%,32.7% and 64.9%,respectively.JC-1 revealed a decrease in Ψ m.Apoptosis of HepG2 cells was associated with caspase-3 and caspase-9 activation,down-regulation of Bcl-2,Mcl-1,and Bcl-xl,and no change in Bax.CONCLUSIONS:Harmine had an anti-proliferative effect in HepG2 cells by inducing apoptosis.Mitochondrial signal pathways were involved in the apoptosis.The cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma.</description><identifier>ISSN: 1499-3872</identifier><identifier>DOI: 10.1016/S1499-3872(11)60102-1</identifier><identifier>PMID: 22146623</identifier><language>eng</language><publisher>Singapore: Elsevier B.V</publisher><subject>apoptosis ; Apoptosis - drug effects ; Apoptosis - genetics ; Bcl-2 ; Bcl-2 protein ; Blotting, Western ; carcinoma ; caspase-3 ; Cell Proliferation - drug effects ; Endocrinology & Metabolism ; Flow Cytometry ; Gastroenterology and Hepatology ; harmine ; Harmine - pharmacology ; Hep G2 Cells ; hepatocellular ; hepatocellular carcinoma ; Humans ; Membrane Potential, Mitochondrial - drug effects ; Microscopy, Fluorescence ; Mitochondria, Liver - drug effects ; Mitochondria, Liver - genetics ; Mitochondria, Liver - metabolism ; Monoamine Oxidase Inhibitors - pharmacology ; protein ; Signal Transduction - drug effects</subject><ispartof>Hepatobiliary & pancreatic diseases international, 2011-12, Vol.10 (6), p.599-604</ispartof><rights>The Editorial Board of Hepatobiliary & Pancreatic Diseases International</rights><rights>2011 The Editorial Board of Hepatobiliary & Pancreatic Diseases International</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c499t-7908e150af6ccffe345784dedbfa19f7ea3fcfac9c08adb2a0c4198bee9526e53</citedby><cites>FETCH-LOGICAL-c499t-7908e150af6ccffe345784dedbfa19f7ea3fcfac9c08adb2a0c4198bee9526e53</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/89801X/89801X.jpg</thumbnail><linktohtml>$$Uhttps://www.sciencedirect.com/science/article/pii/S1499387211601021$$EHTML$$P50$$Gelsevier$$H</linktohtml><link.rule.ids>314,776,780,3537,27903,27904,65309</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/22146623$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Cao, Ming-Rong</creatorcontrib><creatorcontrib>Li, Qiang</creatorcontrib><creatorcontrib>Liu, Zhi-Long</creatorcontrib><creatorcontrib>Liu, Hui-Hui</creatorcontrib><creatorcontrib>Wang, Wei</creatorcontrib><creatorcontrib>Liao, Xiao-Li</creatorcontrib><creatorcontrib>Pan, Yun-Long</creatorcontrib><creatorcontrib>Jiang, Jian-Wei</creatorcontrib><title>Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway</title><title>Hepatobiliary & pancreatic diseases international</title><addtitle>Hepatobiliary & Pancreatic Diseases International</addtitle><description>BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA topoisomerase.However no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma.This study aimed to investigate the effects of harmine on proliferation and apoptosis and the underlying mechanisms in the human hepatocellular carcinoma cell line HepG2.METHODS:The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation test.The morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell cycle.Western blotting was used to assess expression of the apoptosis-regulated genes Bcl-2,Bax,Bcl-xl,Mcl-1,caspase-3,and caspase-9 Mitochondrial transmembrane potential (Ψ m) was determined using JC-1.RESULTS:Harmine inhibited the proliferation of HepG2 cells in a dose-dependent manner.Hoechst 33258 staining revealed nuclear fragmentation and chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with harmine.The percentage of the sub/G1 fraction was increased in a concentration-dependent manner,indicating apoptotic cell death.PI staining showed that harmine changed the cell cycle distribution,by decreasing the proportion of cells inG0/G1 and increasing the proportion in S and G2/M.Harmine induced apoptosis in a concentration-dependent manner,with rates of 20.0%,32.7% and 64.9%,respectively.JC-1 revealed a decrease in Ψ m.Apoptosis of HepG2 cells was associated with caspase-3 and caspase-9 activation,down-regulation of Bcl-2,Mcl-1,and Bcl-xl,and no change in Bax.CONCLUSIONS:Harmine had an anti-proliferative effect in HepG2 cells by inducing apoptosis.Mitochondrial signal pathways were involved in the apoptosis.The cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma.</description><subject>apoptosis</subject><subject>Apoptosis - drug effects</subject><subject>Apoptosis - genetics</subject><subject>Bcl-2</subject><subject>Bcl-2 protein</subject><subject>Blotting, Western</subject><subject>carcinoma</subject><subject>caspase-3</subject><subject>Cell Proliferation - drug effects</subject><subject>Endocrinology & Metabolism</subject><subject>Flow Cytometry</subject><subject>Gastroenterology and Hepatology</subject><subject>harmine</subject><subject>Harmine - pharmacology</subject><subject>Hep G2 Cells</subject><subject>hepatocellular</subject><subject>hepatocellular carcinoma</subject><subject>Humans</subject><subject>Membrane Potential, Mitochondrial - drug effects</subject><subject>Microscopy, Fluorescence</subject><subject>Mitochondria, Liver - drug effects</subject><subject>Mitochondria, Liver - genetics</subject><subject>Mitochondria, Liver - metabolism</subject><subject>Monoamine Oxidase Inhibitors - pharmacology</subject><subject>protein</subject><subject>Signal Transduction - drug effects</subject><issn>1499-3872</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNqFkU9v1DAQxX0AtaXlI4AicQAOAY-df75QoQq6SCv1AJytWWe8dUni1E6K9tvj7C4V4tLTSKM3b-b9hrFXwD8Ah-rjdyiUymVTi3cA7ysOXOTwjJ09tk_ZixjvOBdNU1Yn7FQIKKpKyDO2XmHo3UCZG9rZUMxw9OPko4upk61ovBaZoa6L2YPDrHeTN7d-aIPDLotuO2Dnhm024nT7G3cX7LnFLtLLYz1nP79--XG1ytc319-uPq9zkw6a8lrxhqDkaCtjrCVZlHVTtNRuLIKyNaG0xqJRhjfYbgRyU4BqNkSqFBWV8py9PfiOwd_PFCfdu7hciQP5OWrFVV0WXC7K8qA0wccYyOoxuB7DTgPXCzu9Z6cXSBpA79lpSHOvjxvmTU_t49RfcElweRBQyvngKOhoHA2GWhfITLr17skVn_5zMImlM9j9oh3FOz-HRDdq0FFofjBZPAD2DovBm2O29JHtffrDP-G45FKoppZ_AO9KohM</recordid><startdate>20111201</startdate><enddate>20111201</enddate><creator>Cao, Ming-Rong</creator><creator>Li, Qiang</creator><creator>Liu, Zhi-Long</creator><creator>Liu, Hui-Hui</creator><creator>Wang, Wei</creator><creator>Liao, Xiao-Li</creator><creator>Pan, Yun-Long</creator><creator>Jiang, Jian-Wei</creator><general>Elsevier B.V</general><scope>2RA</scope><scope>92L</scope><scope>CQIGP</scope><scope>W91</scope><scope>~WA</scope><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>20111201</creationdate><title>Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway</title><author>Cao, Ming-Rong ; Li, Qiang ; Liu, Zhi-Long ; Liu, Hui-Hui ; Wang, Wei ; Liao, Xiao-Li ; Pan, Yun-Long ; Jiang, Jian-Wei</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c499t-7908e150af6ccffe345784dedbfa19f7ea3fcfac9c08adb2a0c4198bee9526e53</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>apoptosis</topic><topic>Apoptosis - drug effects</topic><topic>Apoptosis - genetics</topic><topic>Bcl-2</topic><topic>Bcl-2 protein</topic><topic>Blotting, Western</topic><topic>carcinoma</topic><topic>caspase-3</topic><topic>Cell Proliferation - drug effects</topic><topic>Endocrinology & Metabolism</topic><topic>Flow Cytometry</topic><topic>Gastroenterology and Hepatology</topic><topic>harmine</topic><topic>Harmine - pharmacology</topic><topic>Hep G2 Cells</topic><topic>hepatocellular</topic><topic>hepatocellular carcinoma</topic><topic>Humans</topic><topic>Membrane Potential, Mitochondrial - drug effects</topic><topic>Microscopy, Fluorescence</topic><topic>Mitochondria, Liver - drug effects</topic><topic>Mitochondria, Liver - genetics</topic><topic>Mitochondria, Liver - metabolism</topic><topic>Monoamine Oxidase Inhibitors - pharmacology</topic><topic>protein</topic><topic>Signal Transduction - drug effects</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Cao, Ming-Rong</creatorcontrib><creatorcontrib>Li, Qiang</creatorcontrib><creatorcontrib>Liu, Zhi-Long</creatorcontrib><creatorcontrib>Liu, Hui-Hui</creatorcontrib><creatorcontrib>Wang, Wei</creatorcontrib><creatorcontrib>Liao, Xiao-Li</creatorcontrib><creatorcontrib>Pan, Yun-Long</creatorcontrib><creatorcontrib>Jiang, Jian-Wei</creatorcontrib><collection>中文科技期刊数据库</collection><collection>中文科技期刊数据库-CALIS站点</collection><collection>中文科技期刊数据库-7.0平台</collection><collection>中文科技期刊数据库-医药卫生</collection><collection>中文科技期刊数据库- 镜像站点</collection><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>Hepatobiliary & pancreatic diseases international</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Cao, Ming-Rong</au><au>Li, Qiang</au><au>Liu, Zhi-Long</au><au>Liu, Hui-Hui</au><au>Wang, Wei</au><au>Liao, Xiao-Li</au><au>Pan, Yun-Long</au><au>Jiang, Jian-Wei</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway</atitle><jtitle>Hepatobiliary & pancreatic diseases international</jtitle><addtitle>Hepatobiliary & Pancreatic Diseases International</addtitle><date>2011-12-01</date><risdate>2011</risdate><volume>10</volume><issue>6</issue><spage>599</spage><epage>604</epage><pages>599-604</pages><issn>1499-3872</issn><abstract>BACKGROUND:Harmine has antitumor and antinociceptive effects,and inhibits human DNA topoisomerase.However no detailed data are available on the mechanisms of action of harmine in hepatocellular carcinoma.This study aimed to investigate the effects of harmine on proliferation and apoptosis and the underlying mechanisms in the human hepatocellular carcinoma cell line HepG2.METHODS:The proliferation of HepG2 cells was determined by the cell counting kit-8 (CCK-8) assay and the clone formation test.The morphology of HepG2 cells was examined using fluorescence microscopy after Hoechst 33258 staining Annexin V/propidium iodide (PI) was used to analyze apoptosis and PI to analyze the cell cycle.Western blotting was used to assess expression of the apoptosis-regulated genes Bcl-2,Bax,Bcl-xl,Mcl-1,caspase-3,and caspase-9 Mitochondrial transmembrane potential (Ψ m) was determined using JC-1.RESULTS:Harmine inhibited the proliferation of HepG2 cells in a dose-dependent manner.Hoechst 33258 staining revealed nuclear fragmentation and chromosomal condensation,cell shrinkage,and attachment loss in HepG2 cells treated with harmine.The percentage of the sub/G1 fraction was increased in a concentration-dependent manner,indicating apoptotic cell death.PI staining showed that harmine changed the cell cycle distribution,by decreasing the proportion of cells inG0/G1 and increasing the proportion in S and G2/M.Harmine induced apoptosis in a concentration-dependent manner,with rates of 20.0%,32.7% and 64.9%,respectively.JC-1 revealed a decrease in Ψ m.Apoptosis of HepG2 cells was associated with caspase-3 and caspase-9 activation,down-regulation of Bcl-2,Mcl-1,and Bcl-xl,and no change in Bax.CONCLUSIONS:Harmine had an anti-proliferative effect in HepG2 cells by inducing apoptosis.Mitochondrial signal pathways were involved in the apoptosis.The cancer-specific selectivity shown in this study suggested that harmine is a promising novel drug for human hepatocellular carcinoma.</abstract><cop>Singapore</cop><pub>Elsevier B.V</pub><pmid>22146623</pmid><doi>10.1016/S1499-3872(11)60102-1</doi><tpages>6</tpages></addata></record> |
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| subjects | apoptosis Apoptosis - drug effects Apoptosis - genetics Bcl-2 Bcl-2 protein Blotting, Western carcinoma caspase-3 Cell Proliferation - drug effects Endocrinology & Metabolism Flow Cytometry Gastroenterology and Hepatology harmine Harmine - pharmacology Hep G2 Cells hepatocellular hepatocellular carcinoma Humans Membrane Potential, Mitochondrial - drug effects Microscopy, Fluorescence Mitochondria, Liver - drug effects Mitochondria, Liver - genetics Mitochondria, Liver - metabolism Monoamine Oxidase Inhibitors - pharmacology protein Signal Transduction - drug effects |
| title | Harmine induces apoptosis in HepG2 cells via mitochondrial signaling pathway |
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