Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells
AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture...
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| Veröffentlicht in: | World journal of gastroenterology : WJG 2008-06, Vol.14 (24), p.3849-3854 |
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| creator | Xin, Xiao-Min Li, Gui-Qiu Jin, Ying-Yu Zhuang, Min Li, Di |
| description | AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay （ELISA）. Intracellular viral DNA and covalently closed circular DNA （cccDNA） were quantified by real-time polymerase chain reaction （PCR）. HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR （RT-PCR）. RESULTS： siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dosedependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantlycombination of siRNAs significantly suppressed HBV cccDNA amplification. CONCLUSION： Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells, especially on cccDNA amplification. |
| doi_str_mv | 10.3748/wjg.14.3849 |
| format | Article |
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METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay （ELISA）. Intracellular viral DNA and covalently closed circular DNA （cccDNA） were quantified by real-time polymerase chain reaction （PCR）. HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR （RT-PCR）. RESULTS： siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dosedependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantlycombination of siRNAs significantly suppressed HBV cccDNA amplification. CONCLUSION： Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells, especially on cccDNA amplification.</description><identifier>ISSN: 1007-9327</identifier><identifier>EISSN: 2219-2840</identifier><identifier>DOI: 10.3748/wjg.14.3849</identifier><identifier>PMID: 18609708</identifier><language>eng</language><publisher>United States: Department of Laboratory Diagnosis,the First Affiliated Hospital of Harbin Medical University,Harbin 150081,Heilongjiang Province,China%Department of Microbiology,Harbin Medical University,Harbin 150081,Heilongjiang Province,China</publisher><subject>Basic Research ; Carcinoma, Hepatocellular - immunology ; Carcinoma, Hepatocellular - metabolism ; Carcinoma, Hepatocellular - virology ; Cell Line, Tumor ; DNA ; DNA, Viral - drug effects ; DNA, Viral - metabolism ; Dose-Response Relationship, Drug ; Hepacivirus - genetics ; Hepacivirus - physiology ; Hepatitis B e Antigens - drug effects ; Hepatitis B e Antigens - metabolism ; Hepatitis B Surface Antigens - drug effects ; Hepatitis B Surface Antigens - metabolism ; Humans ; Liver Neoplasms - immunology ; Liver Neoplasms - metabolism ; Liver Neoplasms - virology ; Nucleic Acid Amplification Techniques ; Plasmids ; RNA, Messenger - metabolism ; RNA, Small Interfering - pharmacology ; RNAs ; RNA干扰 ; Transfection ; Virus Replication - drug effects ; 乙肝病毒</subject><ispartof>World journal of gastroenterology : WJG, 2008-06, Vol.14 (24), p.3849-3854</ispartof><rights>Copyright © Wanfang Data Co. Ltd. All Rights Reserved.</rights><rights>2008 The WJG Press and Baishideng. All rights reserved. 2008</rights><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c435t-9d42290c622b13d4fb8402d7bdb3fda55ca63ab5351adc1faeed4afed272a9703</citedby><cites>FETCH-LOGICAL-c435t-9d42290c622b13d4fb8402d7bdb3fda55ca63ab5351adc1faeed4afed272a9703</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Uhttp://image.cqvip.com/vip1000/qk/84123X/84123X.jpg</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2721441/pdf/$$EPDF$$P50$$Gpubmedcentral$$H</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2721441/$$EHTML$$P50$$Gpubmedcentral$$H</linktohtml><link.rule.ids>230,314,723,776,780,881,27901,27902,53766,53768</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/18609708$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Xin, Xiao-Min</creatorcontrib><creatorcontrib>Li, Gui-Qiu</creatorcontrib><creatorcontrib>Jin, Ying-Yu</creatorcontrib><creatorcontrib>Zhuang, Min</creatorcontrib><creatorcontrib>Li, Di</creatorcontrib><title>Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells</title><title>World journal of gastroenterology : WJG</title><addtitle>World Journal of Gastroenterology</addtitle><description>AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay （ELISA）. Intracellular viral DNA and covalently closed circular DNA （cccDNA） were quantified by real-time polymerase chain reaction （PCR）. HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR （RT-PCR）. RESULTS： siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dosedependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantlycombination of siRNAs significantly suppressed HBV cccDNA amplification. CONCLUSION： Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells, especially on cccDNA amplification.</description><subject>Basic Research</subject><subject>Carcinoma, Hepatocellular - immunology</subject><subject>Carcinoma, Hepatocellular - metabolism</subject><subject>Carcinoma, Hepatocellular - virology</subject><subject>Cell Line, Tumor</subject><subject>DNA</subject><subject>DNA, Viral - drug effects</subject><subject>DNA, Viral - metabolism</subject><subject>Dose-Response Relationship, Drug</subject><subject>Hepacivirus - genetics</subject><subject>Hepacivirus - physiology</subject><subject>Hepatitis B e Antigens - drug effects</subject><subject>Hepatitis B e Antigens - metabolism</subject><subject>Hepatitis B Surface Antigens - drug effects</subject><subject>Hepatitis B Surface Antigens - metabolism</subject><subject>Humans</subject><subject>Liver Neoplasms - immunology</subject><subject>Liver Neoplasms - metabolism</subject><subject>Liver Neoplasms - virology</subject><subject>Nucleic Acid Amplification Techniques</subject><subject>Plasmids</subject><subject>RNA, Messenger - metabolism</subject><subject>RNA, Small Interfering - pharmacology</subject><subject>RNAs</subject><subject>RNA干扰</subject><subject>Transfection</subject><subject>Virus Replication - drug effects</subject><subject>乙肝病毒</subject><issn>1007-9327</issn><issn>2219-2840</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2008</creationdate><recordtype>article</recordtype><sourceid>EIF</sourceid><recordid>eNpVkc1v1DAQxS0EokvhxB1ZiBtKao-dTXJBWhZoK1VFQnC2Jv7Iesk6wc626n-Pl11ROM1h3vzmzTxCXnNWilo2F_fbvuSyFI1sn5AFAG8LaCR7ShacsbpoBdRn5EVKW8ZAiAqekzPeLFlbs2ZB4nrcdT7g7MdAR0fTDoeB-jDb6Gz0oaffbleJ7qzxONtE-2hzjTTtpynalP6MBbqxU0bMPtGP9M7HfaJa60-3q0yiV3a6hBJKXlFthyG9JM8cDsm-OtVz8uPL5-_rq-Lm6-X1enVTaCmquWiNBGiZXgJ0XBjpunwUmLoznXAGq0rjUmBXiYqj0dyhtUaiswZqwHycOCcfjtxp32X_2oY54qCm6HcYH9SIXv3fCX6j-vFOZQCXkmfAuyPgHoPD0KvtuI8hW1b55cBYA5Lxw573R5mOY0rRur8rOFOHhA5yxaU6JJTVb_519ag9RZIFb0-4zRj6XzkC1aH-6fxgs7NGyqquxW99Epl9</recordid><startdate>20080628</startdate><enddate>20080628</enddate><creator>Xin, Xiao-Min</creator><creator>Li, Gui-Qiu</creator><creator>Jin, Ying-Yu</creator><creator>Zhuang, Min</creator><creator>Li, Di</creator><general>Department of Laboratory Diagnosis,the First Affiliated Hospital of Harbin Medical University,Harbin 150081,Heilongjiang Province,China%Department of Microbiology,Harbin Medical University,Harbin 150081,Heilongjiang Province,China</general><general>The WJG Press and Baishideng</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>2B.</scope><scope>4A8</scope><scope>92I</scope><scope>93N</scope><scope>PSX</scope><scope>TCJ</scope><scope>5PM</scope></search><sort><creationdate>20080628</creationdate><title>Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells</title><author>Xin, Xiao-Min ; Li, Gui-Qiu ; Jin, Ying-Yu ; Zhuang, Min ; Li, Di</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c435t-9d42290c622b13d4fb8402d7bdb3fda55ca63ab5351adc1faeed4afed272a9703</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2008</creationdate><topic>Basic Research</topic><topic>Carcinoma, Hepatocellular - immunology</topic><topic>Carcinoma, Hepatocellular - metabolism</topic><topic>Carcinoma, Hepatocellular - virology</topic><topic>Cell Line, Tumor</topic><topic>DNA</topic><topic>DNA, Viral - drug effects</topic><topic>DNA, Viral - metabolism</topic><topic>Dose-Response Relationship, Drug</topic><topic>Hepacivirus - genetics</topic><topic>Hepacivirus - physiology</topic><topic>Hepatitis B e Antigens - drug effects</topic><topic>Hepatitis B e Antigens - metabolism</topic><topic>Hepatitis B Surface Antigens - drug effects</topic><topic>Hepatitis B Surface Antigens - metabolism</topic><topic>Humans</topic><topic>Liver Neoplasms - immunology</topic><topic>Liver Neoplasms - metabolism</topic><topic>Liver Neoplasms - virology</topic><topic>Nucleic Acid Amplification Techniques</topic><topic>Plasmids</topic><topic>RNA, Messenger - metabolism</topic><topic>RNA, Small Interfering - pharmacology</topic><topic>RNAs</topic><topic>RNA干扰</topic><topic>Transfection</topic><topic>Virus Replication - drug effects</topic><topic>乙肝病毒</topic><toplevel>online_resources</toplevel><creatorcontrib>Xin, Xiao-Min</creatorcontrib><creatorcontrib>Li, Gui-Qiu</creatorcontrib><creatorcontrib>Jin, Ying-Yu</creatorcontrib><creatorcontrib>Zhuang, Min</creatorcontrib><creatorcontrib>Li, Di</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>Wanfang Data Journals - Hong Kong</collection><collection>WANFANG Data Centre</collection><collection>Wanfang Data Journals</collection><collection>万方数据期刊 - 香港版</collection><collection>China Online Journals (COJ)</collection><collection>China Online Journals (COJ)</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>World journal of gastroenterology : WJG</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Xin, Xiao-Min</au><au>Li, Gui-Qiu</au><au>Jin, Ying-Yu</au><au>Zhuang, Min</au><au>Li, Di</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells</atitle><jtitle>World journal of gastroenterology : WJG</jtitle><addtitle>World Journal of Gastroenterology</addtitle><date>2008-06-28</date><risdate>2008</risdate><volume>14</volume><issue>24</issue><spage>3849</spage><epage>3854</epage><pages>3849-3854</pages><issn>1007-9327</issn><eissn>2219-2840</eissn><abstract>AIM： To observe the inhibition of hepatitis B virus （HBV） replication and expression in HepG2.2.15 cells by combination of small interfering RNAs （siRNAs）. METHODS： Recombinant plasmid psiI-HBV was constructed and transfected into HepG2.2.15 cells. At 48 h, 72 h and 96 h after transfection, culture media were collected and cells were harvested for HBV replication assay. HBsAg and HBeAg in the cell culture medium were detected by enzyme-linked immunoadsorbent assay （ELISA）. Intracellular viral DNA and covalently closed circular DNA （cccDNA） were quantified by real-time polymerase chain reaction （PCR）. HBV viral mRNA was reverse transcribed and quantified by reverse-transcript PCR （RT-PCR）. RESULTS： siRNAs showed marked anti-HBV effects. siRNAs could specifically inhibit the expression of HBsAg and the replication of HBV DNA in a dosedependent manner. Furthermore, combination of siRNAs, compared with individual use of each siRNA, exerted a stronger inhibition on antigen expression and viral replication. More importantlycombination of siRNAs significantly suppressed HBV cccDNA amplification. CONCLUSION： Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells, especially on cccDNA amplification.</abstract><cop>United States</cop><pub>Department of Laboratory Diagnosis,the First Affiliated Hospital of Harbin Medical University,Harbin 150081,Heilongjiang Province,China%Department of Microbiology,Harbin Medical University,Harbin 150081,Heilongjiang Province,China</pub><pmid>18609708</pmid><doi>10.3748/wjg.14.3849</doi><tpages>6</tpages><oa>free_for_read</oa></addata></record> |
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| source | MEDLINE; Baishideng "World Journal of" online journals; EZB-FREE-00999 freely available EZB journals; PubMed Central; Alma/SFX Local Collection |
| subjects | Basic Research Carcinoma, Hepatocellular - immunology Carcinoma, Hepatocellular - metabolism Carcinoma, Hepatocellular - virology Cell Line, Tumor DNA DNA, Viral - drug effects DNA, Viral - metabolism Dose-Response Relationship, Drug Hepacivirus - genetics Hepacivirus - physiology Hepatitis B e Antigens - drug effects Hepatitis B e Antigens - metabolism Hepatitis B Surface Antigens - drug effects Hepatitis B Surface Antigens - metabolism Humans Liver Neoplasms - immunology Liver Neoplasms - metabolism Liver Neoplasms - virology Nucleic Acid Amplification Techniques Plasmids RNA, Messenger - metabolism RNA, Small Interfering - pharmacology RNAs RNA干扰 Transfection Virus Replication - drug effects 乙肝病毒 |
| title | Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells |
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