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

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 bec...

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Autores principales: Shier, Medhat K., El-Wetidy, Mohammad S., Ali, Hebatallah H., Al-Qattan, Mohammad M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Medknow Publications & Media Pvt Ltd 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4898095/
https://www.ncbi.nlm.nih.gov/pubmed/27184644
http://dx.doi.org/10.4103/1319-3767.182461
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author Shier, Medhat K.
El-Wetidy, Mohammad S.
Ali, Hebatallah H.
Al-Qattan, Mohammad M.
author_facet Shier, Medhat K.
El-Wetidy, Mohammad S.
Ali, Hebatallah H.
Al-Qattan, Mohammad M.
author_sort Shier, Medhat K.
collection PubMed
description 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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spelling pubmed-48980952016-06-13 Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A Shier, Medhat K. El-Wetidy, Mohammad S. Ali, Hebatallah H. Al-Qattan, Mohammad M. Saudi J Gastroenterol Original Article 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. Medknow Publications & Media Pvt Ltd 2016 /pmc/articles/PMC4898095/ /pubmed/27184644 http://dx.doi.org/10.4103/1319-3767.182461 Text en Copyright: © 2016 Saudi Journal of Gastroenterology (Official journal of The Saudi Gastroenterology Association) http://creativecommons.org/licenses/by-nc-sa/3.0 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.
spellingShingle Original Article
Shier, Medhat K.
El-Wetidy, Mohammad S.
Ali, Hebatallah H.
Al-Qattan, Mohammad M.
Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A
title Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A
title_full Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A
title_fullStr Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A
title_full_unstemmed Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A
title_short Hepatitis C Virus Genotype 4 Replication in the Hepatocellular Carcinoma Cell Line HepG2/C3A
title_sort hepatitis c virus genotype 4 replication in the hepatocellular carcinoma cell line hepg2/c3a
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4898095/
https://www.ncbi.nlm.nih.gov/pubmed/27184644
http://dx.doi.org/10.4103/1319-3767.182461
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