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Combination of small interfering RNAs mediates greater suppression on hepatitis B virus cccDNA in HepG2.2.15 cells

查看全文 作　　者：Xiao-Min [1]Xin;Gui-Qiu [1]Li;Ying-Yu [1]Jin;Min [2]Zhuang;Di [2]Li 高影响力作者 机构地区：[1]Department ofLaboratory Diagnosis, the First Affiliated Hospital of HarbinMedical University, Harbin 150081, Heilongjiang Province,China;[2]Department of Microbiology, HarbinMedical University, Harbin 150081, Heilongjiang Province,China高影响力机构 出　　处：《World Journal of Gastroenterology》索引2008年第14卷第24期,共6页高影响力期刊 基　　金：The Youth Foundation of Heilongjiang Province,No.QC06C061;the Foundation of Education Department,Heilongjiang Province,No.11521089 摘　　要：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 psil-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 quantifi ed by real-time polymerase chain reaction (PCR). HBV viral mRNA was reverse transcribed and quantifi ed 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 dose-dependent manner. Furthermore,combination of siRNAs,compared with individual use of each siRNA,exerted a stronger inhibition on antigen expression and viral replication. More importantly,combination of siRNAs significantly suppressed HBV cccDNA amplifi cation. CONCLUSION: Combination of siRNAs mediates a stronger inhibition on viral replication and antigenexpression in HepG2.2.15 cells,especially on cccDNA amplifi cation. 关 键 词：RNAS DNA 乙肝病毒 RNA干扰
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