|Table of Contents|

Effect of miRNA-146a on life cycle of hepatitis B virus in vitro



Research Field:
Publishing date:



Effect of miRNA-146a on life cycle of hepatitis B virus in vitro


ZHAO Min ZHANG Zhenzhen LIU Quanbo

Department of Infectious Disease, Key Laboratory of Child Development and Disorders of Ministry of Education, International Science and Technology Cooperation Base of Child Development and Disorders, Chongqing Key Laboratory of Pediatrics, Children’s Hospital of Chongqing Medical University, Chongqing, 400014, China


hepatitis B virus miR-146a heparan sulfate-glucosamine 3-sulfotransferase 3

R373.21; R39433; R394.3

Objective     To determine the effect of microRNA-146a (miR-146a) on the life cycle of hepatitis B virus (HBV) and investigate the underlying mechanisms. Methods     The miRNA expression profiles were compared by miRNA array between HepG2 and HepG2.2.15 cells. Then miR-146a was chosen as objective, and its expression level was further confirmed by RT-PCR. After miR-146a mimic and inhibitor were transfected into HepG2.2.15 cells respectively, the quantification of HBV replication was determined by RT-PCR, and the levels of HBsAg and HBeAg in the supernatant were measured by ELISA, and the expression of HS3ST3B1 at mRNA and protein levels were tested by RT-PCR and Western blotting.  Dual-luciferase reporter assay was used to detect the interaction between miR-146a and potential target HS3ST3B1. Results     The expression levels of totally 72 miRNAs were changed in HepG2.2.15 cells, with 27 up-regulated and 45 down-regulated. RT-PCR showed the expression level of miR-146a was significantly higher in HepG2.2.15 cells than HepG2 cells (1.55±0.13 vs 1.00±0.01, P<0.05). Transfection of miR-146a mimic into HepG2.2.15 cells resulted in significantly increased HBV replication and levels of HBsAg and HBeAg (P<0.05), while the transfection of its inhibited caused opposite results (P<0.05).Bioinformatic analysis showed that HS3ST3B1 was a potential target of miR-146a. The reporter luciferase reporter system indicated that the reported fluorescence intensity of HS3ST3B1 wild type vector was significantly lower than that of the control group (P<0.05), but showed no significant difference between HS3ST3B1 mutant vector and control group (P>0.05). The mRNA level of HS3ST3B1 was not significantly changed in HepG2.2.15 cells transfected with miR-146a mimic (P>0.05), but its protein level was significantly decreased (P<0.05).  Conclusions    miR-146a affects the life cycle of HBV, which  may be through suppressing the translation of HBV inhibitory factor HS3ST3B1 3′UTR.


[1]LAMONTAGNE R J, BAGGA S, BOUCHARD M J. Hepatitis B virus molecular biology and pathogenesis[J]. Hepatoma Res, 2016, 2: 163-186. DOI:10.20517/2394-5079.2016.05.
[2]ZHANG C, ZHONG Y, GUO L. Strategies to prevent hepatitis B virus infection in China: immunization, screening, and standard medical practices[J]. Biosci Trends, 2013, 7(1): 7-12.
[3]HOOFNAGLE J H, LAU D. New therapies for chronic hepatitis B[J]. J Viral Hepat, 1997, 4(Suppl 1): 41-50. DOI:10.1111/liv.13086.
[4]EL-SERAG H B. Epidemiology of viral hepatitis and hepatocellular carcinoma[J]. Gastroenterology, 2012, 142(6): 1264-1273.e1. DOI:10.1053/j.gastro.2011.12.061.
[5]BLUM H E. History and Global Burden of Viral Hepatitis[J]. Dig Dis, 2016, 34(4): 293-302. DOI:10.1159/000444466.
[6]ZOULIM F, DURANTEL D. Antiviral Therapies and Prospects for a Cure of Chronic Hepatitis B[J]. Cold Spring Harb Perspect Med, 2015, 5(4). DOI:10.1101/cshperspect.a021501.
[7]GISH R G, GIVEN B D, LAI C L, et al. Chronic hepatitis B: Virology, natural history, current management and a glimpse at future opportunities[J]. Antiviral Res, 2015, 121: 47-58. DOI:10.1016/j.antiviral.2015.06.008.
[8]LIi L, CHEN X P, LI Y J. MicroRNA-146a and human disease[J]. Scand J Immunol, 2010, 71(4): 227-231. DOI:10.1111/j. 1365-3083.2010.02383.x.
[9]SABA R, SORENSEN D L, BOOTH S A. MicroRNA146a: a dominant, negative regulator of the innate immune response[J]. Front Immunol, 2014, 5:578. DOI: 10.3389/fimmu.2014.00578.
[10]HAYES C N, CHAYAMA K. MicroRNAs as biomarkers for liver disease and hepatocellular carcinoma[J]. Int J Mol Sci, 2016, 17(3):280. DOI: 10.3390/ijms17030280.
[11]REN J P, YING R S, CHENG Y Q, et al. HCV-induced miR146a controls SOCS1/STAT3 and cytokine expression in monocytes to promote regulatory T-cell development[J]. J Viral Hepat, 2016, 23(10): 755-766. DOI:10.1111/jvh.12537.
[12]RUSCA N, MONTICELLI S. MiR-146a in Immunity and Disease[J]. Mol Biol Int, 2011, 2011: 437301. DOI:10.4061/2011/437301.
[13]ZHAO J L, STARCZYNOWSKI D T. Role of microRNA-146a in normal and malignant hematopoietic stem cell function[J]. Front Genet, 2014, 9; 5:219. DOI: 10.3389/fgene.2014.00219.eCollection 2014.
[14]SELLS M A, ZELENT A Z, SHVARTSMAN M, et al. Replicative intermediates of hepatitis B virus in HepG2 cells that produce infectious virions[J]. J Virol, 1988, 62(8): 2836-2844.
[15]ZHANG Z, LIU X, CHEN J, et al. Heparin sulphate D-glucosaminyl 3-O-sulfotransferase 3B1 plays a role in HBV replication[J]. Virology, 2010, 406(2): 280-285. DOI:10. 1016/j.virol.2010.07.030.
[16]HOU Z H, HAN Q J, ZHANG C, et al. miR146a impairs the IFN-induced anti-HBV immune response by downregulating STAT1 in hepatocytes[J]. Liver Int, 2014, 34(1): 58-68. DOI:10.1111/liv.12244.
[17]RIAZALHOSSEINI B, MOHAMED Z, APALASAMY Y D, et al. Association between microRNA196A2 and microRNA-146A polymorphisms and progression to cirrhosis and hepatocellular carcinoma in patients with viral hepatitis B[J]. Pharmacogenet Genomics, 2016, 26(2): 74-79. DOI:10.1097/FPC.0000000000000187.
[18]GUO L, YANG X, DUAN T. Altered microRNA expression profile in maternal and fetal liver of HBV transgenic mouse model[J]. J Matern Fetal Neonatal Med, 2012, 25(10): 2071-2077. DOI:10.3109/14767058.2012.678431.
[19]WANG S, ZHANG X, JU Y, et al. MicroRNA-146a feedback suppresses T cell immune function by targeting Stat1 in patients with chronic hepatitis B[J]. J Immunol, 2013, 191(1): 293-301. DOI:10.4049/jimmunol.1202100.
[20]HOU Z, ZHANG J, HAN Q, et al. Hepatitis B virus inhibits intrinsic RIG-I and RIG-G immune signaling via inducing miR146a[J]. Sci Rep, 2016, 6:26150.DOI: 10.1038/srep26150.
[21]LINDOW M, KAUPPINEN S. Discovering the first microRNA-targeted drug[J]. J Cell Biol, 2012, 199(3): 407-412. DOI:10.1083/jcb.201208082.


Last Update: 2017-09-04