Huh7.5.1Cells Infected With HCV And Interferon Treatment Process Mi-RNA Expression In The Changes Of The Spectrum

Hepatitis C is a liver as the main target organ, infectious disease caused by the hepatitis C virus (HCV). According to the World Health Organization, the global hepatitis C virus infection in patients of about170million people, cirrhosis and HCV-related hepatocellular carcinoma (HCC) is the leading cause of death in patients with hepatitis C, resulted in a very life and safety of the people of the world major threat, was actively searching for the way of effective treatment of HCV is crucial.The past10years, the treatment of hepatitis C has made considerable progress, interferon has played an increasingly important role in their treatment process. The development of α-interferon (IFN-a) treatment from the initial single to in recent years promoting the use of pegylated-of IFN-α (the Pegylated Interferon-α, PEG-of IFN-α) plus ribavirin to IFN-α and ribavirin combination therapy; treatment of persistent viral response rate has been greatly improved. However, the treatment effect of alpha-interferon in different people. The function of the realization of α-interferon signaling pathway has been considerable depth, Previous studies have demonstrated that the JAK/STAT cascade signaling α-interferon to reflect its activity, the signal transduction pathway is tightly regulated by multiple mechanisms, in which inhibitory signaling molecules such as SOCS family, can negatively regulate a-interferon signal transduction. In the final α-interferon signal transduction has changed hundreds of gene expression in the cells, By these regulatory genes, alpha-interferon regulation of cell function.However, the difference of the a-interferon treatment of different groups, yet the clear explanation of its mechanism.Gene silencing is an important means of the eukaryotic cell gene expression and regulation. Gene silencing generally occurs on two levels, one is caused by DNA methylation, heterochromatin qualitative and location effects of gene silencing on the transcriptional level (transcriptional gene silencing TGS), Another post-transcriptional gene silencing (post-transcriptional gene silencing, PTGS), that the level of gene transcription through specific degradation of the target RNA and result in gene inactivation. This is to protect their own defense and protection mechanisms, plants and animals in the long-term evolutionary process, the formation of a limit invasion by exogenous nucleic acids.MicroRNA (MiRNA) cells express the rich, highly conserved non-coding small RNA, the length of18-25nt. MiRNA showed a new mechanism for regulation of gene expression in a transcription by binding of target gene mRNA,3’UTR of inhibition of gene expression.The MiRNA has been shown to play a role in inflammation and tumorigenesis in a variety of physiological and pathological processes in the development of infection, the immune response. From initial discovery to the present, has been found in mammals, over700MiRNA, which most of the MiRNA biological function is unknown. Have been reported, the MiRNA involved in the multifaceted role in regulating the immune response. But so far, has not been reported to the MiRNA can regulate interferon signaling pathway thereby affecting the antiviral innate immunity, also did not report the MiRNA involved in interferon regulation of immune cell function.It is based on the above background, we may be in order to explore the hepatitis C virus (HCV) transfection, copy and remove the role of MiRNA, We first passed pFL-J6JFH-1plasmid in vitro transcription to get HCV RNA transcripts, then when Huh7.5.1cells were cultured to reach60%to70%confluence, the HCV RNA transfected into.3days after transfection, extract infected with cell total RNA was amplified by reverse transcription kit, compared with normal in Huh7.5.1cells to determine transfection success. Then deal these infected cells with IFNa-2B, after48h, the re-use of quantitative PCR for detection of intracellular RNA, and without any treatment of the infected cells for comparison. Finally, the MiRNA expression microarray analysis of normal in Huh7.5.1cells of HCV infection in Huh7.5.1cells, and infection after interferon treatment of Huh7.5.1cells were the MiRNA Determination of expression and the MiRNA expression analysis between the two groups of the2(AACT) method.After screening, we found that compared to the MiRNA expression in normal Huh7.5.1cells, There are13kinds of MiRNA in HCV infection upregulated after interferon treatment, a decline; In addition there are seven kinds of MiRNA in HCV infection downregulated after interferon treatment raised. Which MiRNA10a may be associated with viral replication and removal process, MiRNA21may be associated with HCV viral invasion led to the host needs to inhibit transcription, MiRNA149may be related to inhibition of viral replication, MiRNA152may be associated with autoimmune tolerance, There MiRNA99b, miRNA200b, MiRNA210, MiRNA616in HCV infection and interferon treatment change significantly, but did not see reported in the viral infection may be some new mechanism.Conclusion:MiRNA10a, MiRNA21, MiRNA149, MiRNA152, MiRNA99b MiRNA200b, MiRNA210, MiRNA616and other MiRNA have significant changes in experiments are likely to HCV replication, to transcription, clear process, we need further experiments to reveal the relationships between them.