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      Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A

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          Abstract

          Background/Aims:

          The lack of a reliable cell culture system allowing persistent in vitro hepatitis C virus (HCV) propagation is still restraining the search for novel antiviral strategies. HepG2 cells transfection with HCV allows for viral replication. However, the replication is weak presumably because of HepG2 lack of miRNA-122, which is essential for viral replication. Other agents such as polyethylene glycol (PEG) and dimethyl sulfoxide (DMSO) have been shown to increase the efficiency of infection with other viruses. This study included comparison of HCV genotype 4 5′UTR and core RNA levels and HCV core protein expression at different time intervals in the absence or presence of PEG and/or DMSO postinfection.

          Materials and Methods:

          We used serum with native HCV particles in infecting HepG2 cells in vitro. HCV replication was assessed by reverse transcriptase polymerase chain reaction for detection of HCV RNA and immunofluorescence and flow cytometry for detection of HCV core protein.

          Results:

          HCV 5′UTR and core RNA expression was evident at different time intervals after viral infection, especially after cells were treated with PEG. HCV core protein was also evident at different time intervals using both immunofluorescence and flow cytometry. PEG, not DMSO, has increased the HCV core protein expression in the treated cells, similar to its effect on viral RNA expression.

          Conclusions:

          These expression profiles suggest that the current model of cultured HepG2 cells allows the study of HCV genotype 4 replication and different stages of the viral life cycle.

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          Most cited references43

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          Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome.

          A random-primed complementary DNA library was constructed from plasma containing the uncharacterized non-A, non-B hepatitis (NANBH) agent and screened with serum from a patient diagnosed with NANBH. A complementary DNA clone was isolated that was shown to encode an antigen associated specifically with NANBH infections. This clone is not derived from host DNA but from an RNA molecule present in NANBH infections that consists of at least 10,000 nucleotides and that is positive-stranded with respect to the encoded NANBH antigen. These data indicate that this clone is derived from the genome of the NANBH agent and are consistent with the agent being similar to the togaviridae or flaviviridae. This molecular approach should be of great value in the isolation and characterization of other unidentified infectious agents.
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            Epidemiology of hepatitis C virus infection.

            Globally, hepatitis C virus (HCV) has infected an estimated 130 million people, most of whom are chronically infected. HCV-infected people serve as a reservoir for transmission to others and are at risk for developing chronic liver disease, cirrhosis, and primary hepatocellular carcinoma (HCC). It has been estimated that HCV accounts for 27% of cirrhosis and 25% of HCC worldwide. HCV infection has likely been endemic in many populations for centuries. However, the wave of increased HCV-related morbidity and mortality that we are now facing is the result of an unprecedented increase in the spread of HCV during the 20th century. Two 20th century events appear to be responsible for this increase; the widespread availability of injectable therapies and the illicit use of injectable drugs.
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              Clinical significance of hepatitis C virus genotypes.

              Nizar Zein (2000)
              On the basis of phylogenetic analysis of nucleotide sequences, multiple genotypes and subtypes of hepatitis C virus (HCV) have been identified. Characterization of these genetic groups is likely to facilitate and contribute to the development of an effective vaccine against infection with HCV. Differences among HCV genotypes in geographic distributions have provided investigators with an epidemiologic marker that can be used to trace the source of HCV infection in a given population. HCV genotype 1 may represent a more aggressive strain and one that is less likely to respond to interferon treatment than HCV genotype 2 or 3. However, these observations require confirmation before HCV genotyping can be used in clinical settings.
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                Author and article information

                Journal
                Saudi J Gastroenterol
                Saudi J Gastroenterol
                SJG
                Saudi Journal of Gastroenterology : Official Journal of the Saudi Gastroenterology Association
                Medknow Publications & Media Pvt Ltd (India )
                1319-3767
                1998-4049
                May-Jun 2016
                : 22
                : 3
                : 240-248
                Affiliations
                [1 ]College of Medicine Research Center, King Saud University, Riyadh, Saudi Arabia
                [2 ]Department of Medical Microbiology and Immunology, College of Medicine, Menofia University, Egypt
                [3 ]Department of Surgery, College of Medicine, King Saud University, Riyadh, Saudi Arabia
                Author notes
                Address for correspondence: Dr. Medhat K. Shier, College of Medicine Research Center, King Saud University, PO Box 2925 (74), Riyadh - 11461, Saudi Arabia. E-mail: mshier3@ 123456gmail.com
                Article
                SJG-22-240
                10.4103/1319-3767.182461
                4898095
                27184644
                eb4f55fe-46fc-4226-bec0-518a7e2c3ba1
                Copyright: © 2016 Saudi Journal of Gastroenterology (Official journal of The Saudi Gastroenterology Association)

                This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 3.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as the author is credited and the new creations are licensed under the identical terms.

                History
                : 19 November 2015
                : 30 January 2016
                Categories
                Original Article

                Gastroenterology & Hepatology
                dimethyl sulfoxide,hepatitis c virus,hepatitis c virus core,hepatitis c virus 5′utr,hepg2,polyethylene glycol

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