Study Of MicroRNA-206 Repressed HCV Proliferation Through The ACACA Lipid Metabolism Signal Pathway

Hepatitis C virus(HCV)is the positive-strand RNA virus with envelope,which belongs to hepatitis viruses of Flaviviridae.HCV is the cause of hepatitis C disease and owns the ability of immune escape from the host.Since HCV was firstly named in 1989,over 180 million persons were infected with HCV.35 thousand of new HCV patients were found every year,and HCV infection became one of the seriously worldwide public health problems.The transmission routes of HCV infection include blood transmission,sexual behavior,and mother to child transmission and so forth.Fewer of HCV patients could clear the virus without any treatment.However,most of HCV patients(about 80%)developed into chronic HCV infection,and a few of chronic HCV patients could develop into hepatic fibrosis,cirrhosis,hepatic cellular cancer(HCC)or other serious hepatic diseases.HCV infection increase the weight of HCV patients,their family,and society.Studies showed that HCV infection and replication were related with proteins,which participate in lipid metabolism.Decoding the relationship between HCV infection and lipid metabolism is necessary for understanding the pathogenic mechanisms of HCV infection.micro RNAs are important factors to regulate gene expression after transcription,which play nonnegligible roles in metabolism,cell growth,apoptosis,and oncogenesis.micro RNA could regulate proliferation,invasion,migration,and apoptosis of tumors.micro RNA-206(miR-206)is one of the hot-spot micro RNAs in recent studies.miR-206 participated in lipid metabolism via downregulation the nuclear receptor subfamily 1 group H member 3(NR1H3 or LXRα)gene,which could decrease the accumulation of lipid droplets.After HCV infection,the progression of viral replication,proliferation,and pathogenicity are dependent on formation of lipo-viro-particles(LVPs).It is unclear whether miR-206 plays important roles in HCV infection and pathogenesis.Thus,we analyzed the relationship between HCV and miR-206 in this study.At first,we constructed lentiviral vector with miR-206 for overexpression of miR-206 in Huh7.5.1 cell-line.Then,the lentiviral vectors with miR-206 were transfected into Huh7.5.1 cells,and the cells strain steadily expression miR-206 were obtained.In order to study the effect of high-lipid on miR-206 and HCV infection,we constructed the cells with miR-206 and high-lipid culturation(DEME with OA medium).After bioinformatic analysis,we collected six genes(ACACA,PEMT,PLCG1,AGPAT1,HADH and ELOVL4 gene),which were all taken part in lipid metabolism,as target genes regulated by miR-206.After comparison of cells with and without miR-206 transfection,expression levels of the ACACA,PEMT and PLCG1 genes were identified as down-regulation by miR-206.The down-regulated ability of miR-206 existed in cells with both normal control(DMEM)and OA mediums.The repression ability for the ACACA and PEMT genes of miR-206 enhanced after HCV infection,and the repression ability for the ACACA and PLCG1 genes of miR-206 enhanced in cells with OA medium.Thus,the ACACA gene was selected for further study.Importantly,we identified that overexpression of miR-206 could repress HCV proliferation in both DEME and OA mediums.We further supposed miR-206 might influence HCV proliferation through ACACA lipid metabolism signal pathway,so we studied the genes in this pathway.Under condition of HCV infection and DMEM or OA mediums,we compared m RNA expressing levels of the genes in the ACACA signal pathway(including ACACA,NR1H3,SREBF1 and FASN genes).The results showed that the expression level of miR-206 increased at 12 hours after transfection under the condition of DEME,then the expression level reached to highest at 48 hours.The m RNA expression levels of the ACACA,NR1H3,SREBF1 and FASN genes decreased at 24 hours after transfection,but only statistical differences were found in the ACACA and NR1H3 genes.At 48 hours after transfection of miR-206,the m RNA expression levels of all four genes in this pathway showed significantly decreased.Excluding the FASN gene,the m RNA expression levels still showed significant decrease at 72 hours after transfection.Under condition of OA medium,the expression level of miR-206 increase at 48 hours after transfection,but no significant difference was identified.While the m RNA expression levels of four genes in the pathway showed no statistical differences between cells with and without miR-206 transfection.These results might be caused by the limitation of miR-206 expression.In this study,we identified miR-206 could repress HCV proliferation through the ACACA lipid metabolism signal pathway.Our results provided theoretical basis for investigating the roles of miR-206 in HCV infection,proliferation,and pathogenesis.Similarly,our results provided foundations for research and development of miR-206 as a novel drug to treat HCV infection.