title: Suppression of hepatitis B virus replication by microRNA-199a-3p and microRNA-210.
abstract: Accumulating evidence suggests that microRNAs (miRNAs) control the replication of both RNA and DNA viruses. In order to determine whether host-encoded miRNAs affect hepatitis B virus (HBV) replication, antisense oligonucleotides (ASOs) of 328 identified human miRNAs were transfected into HepG2 2.2.15 cells, respectively. ELISA and MTS assay were used to measure the expression level of HBV S protein (HBsAg), HBV e antigen (HBeAg) and cell proliferation. Compared to experimental controls, miR-199a-3p and miR-210 efficiently reduced HBsAg expression without affecting HepG2 2.2.15 cell proliferation. Quantification of HBV DNA by real-time PCR showed that both miRNAs suppressed viral replication. Bioinformatics analysis indicated a putative binding site for miR-199a-3p in the HBsAg coding region and a putative binding site for miR-210 in the HBV pre-S1 region. The direct effect of miRNAs on the target region in HBV transcripts was validated by a fluorescent reporter assay, and the suppression of HBs gene expression by both miRNAs was measured by real-time PCR and Western blot. These results suggest that up-regulation of miR-199a-3p and miR-210 in HepG2 2.2.15 cells compared to HepG2 cells may play a role in regulating HBV replication and maintenance of a suitable level of virion production in persistent infection by targeting crucial HBV genes.
authors: Tianjin Life Science Research Center and Basic Medical School, Tianjin Medical University, Tianjin 300070, China.
authors: Guang-ling
authors: GL
authors: Zhang
authors: Yi-xuan
authors: YX
authors: Li
authors: Shu-qi
authors: SQ
authors: Zheng
authors: Min
authors: M
authors: Liu
authors: Xin
authors: X
authors: Li
authors: Hua
authors: H
authors: Tang
mesh_terms: D000998:Antiviral Agents / Q000494:pharmacology; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D004261:DNA Replication / Q000187:drug effects; D004279:DNA, Viral / Q000096:biosynthesis; D004797:Enzyme-Linked Immunosorbent Assay; D020869:Gene Expression Profiling; D015967:Gene Expression Regulation, Viral / Q000187:drug effects; D057809:HEK293 Cells; D056945:Hep G2 Cells; D006514:Hepatitis B Surface Antigens / Q000096:biosynthesis*; D006513:Hepatitis B e Antigens / Q000096:biosynthesis; D006515:Hepatitis B virus / Q000187:drug effects / Q000235:genetics / Q000502:physiology*; D006801:Humans; D035683:MicroRNAs / Q000502:physiology*; D016133:Polymerase Chain Reaction; D016372:RNA, Antisense / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D014779:Virus Replication / Q000187:drug effects
chemical_list: D000998:Antiviral Agents; D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens; D006513:Hepatitis B e Antigens; C543800:MIRN210 microRNA, human; D035683:MicroRNAs; D016372:RNA, Antisense; D034741:RNA, Small Interfering; C531408:mirn199 microRNA, human