Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus

Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus

The Hepatitis C Virus (HCV) encodes the structural protein Core, which in addition to being the capsid subunit, participates in different mechanisms of HCV infection pathogenesis. Since HCV in vitroreplication system has limitations, the use of viral vectors could be a useful tool to study the Core protein properties. To validate the Semliki Forest Virus (SFV) strategy in transduced HepG2 cells to study the HCV Core protein, the expression of green fluorescent protein (GFP) and Core protein expressions were detected 24 to 96 hours post-transduction in HepG2 cells transduced with rSFV. Core protein expression was lower than GFP expression in HepG2 cells. Since HCV Core protein can regulate the activity of the p53 gene, the transcriptional level of this gene was evaluated. A decrease in the level of p53 mRNA was observed in the cells after transduction, compared to the control cells. Although the cells transduced with rSFV-Core had the lowest level of p53 mRNA, the difference was not significant compared to cells transduced with rSFV-GFP. The results confirm that rSFV allows the transient expression of heterologous proteins in human hepatoma cell lines. Additional studies are needed to determine whether the decreased expression of Core may be due to the degradation of the viral protein. 

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Main Authors: Mira, Catalina, Yepes, Jesús Orlando, Henao, Luis Felipe, Montoya Guzmán, Melissa, Navas, Maria-Cristina
Format: Digital revista
Language:spa
Published: Universidad Nacional de Colombia - Sede Bogotá - Facultad de Ciencias - Departamento de Biología 2020
Online Access:https://revistas.unal.edu.co/index.php/actabiol/article/view/79365
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spelling oai:www.revistas.unal.edu.co:article-793652021-03-23T19:53:27Z Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus EXPRESIÓN DE LA PROTEÍNA CORE DEL VIRUS DE LA HEPATITIS C EN CÉLULAS HEPG2 USANDO EL VIRUS DEL BOSQUE DE SEMLIKI Mira, Catalina Yepes, Jesús Orlando Henao, Luis Felipe Montoya Guzmán, Melissa Navas, Maria-Cristina Expresión transitoria p53 Vector Viral VHC Transitory expression p53 Viral Vector HCV The Hepatitis C Virus (HCV) encodes the structural protein Core, which in addition to being the capsid subunit, participates in different mechanisms of HCV infection pathogenesis. Since HCV in vitroreplication system has limitations, the use of viral vectors could be a useful tool to study the Core protein properties. To validate the Semliki Forest Virus (SFV) strategy in transduced HepG2 cells to study the HCV Core protein, the expression of green fluorescent protein (GFP) and Core protein expressions were detected 24 to 96 hours post-transduction in HepG2 cells transduced with rSFV. Core protein expression was lower than GFP expression in HepG2 cells. Since HCV Core protein can regulate the activity of the p53 gene, the transcriptional level of this gene was evaluated. A decrease in the level of p53 mRNA was observed in the cells after transduction, compared to the control cells. Although the cells transduced with rSFV-Core had the lowest level of p53 mRNA, the difference was not significant compared to cells transduced with rSFV-GFP. The results confirm that rSFV allows the transient expression of heterologous proteins in human hepatoma cell lines. Additional studies are needed to determine whether the decreased expression of Core may be due to the degradation of the viral protein.  El Virus de la Hepatitis C (VHC) codifica la proteína Core. Core, además de ser la subunidad de la cápside, participa en diferentes mecanismos de patogénesis de la infección por VHC. Dado que el sistema de replicación in vitrodel VHC presenta limitaciones, el uso de vectores virales podría ser una herramienta útil para estudiar las propiedades de la proteína Core. Con el fin de validar el vector con el Virus del Bosque de Semliki (SFV) para el estudio de Core en células HepG2, se evaluó la expresión de la proteína verde fluorescente (GFP) y la proteína Core utilizando este vector viral. Las expresiones de GFP y Core se detectaron en células HepG2 transducidas con rSFV de 24 a 96 horas postransducción. La expresión de la proteína Core fue inferior a la expresión de GFP en las células HepG2. Teniendo en cuenta que la proteína Core del VHC puede regular la actividad del gen p53, se evaluó el nivel transcripcional de este gen. Se observó una disminución en el nivel de mARN de p53 en las células luego de la transducción, comparado con las células control. Aunque las células transducidas con rSFV-Core presentaron el menor nivel de mARN de p53,la diferencia no fue significativa comparada con las células transducidas con rSFV-GFP. Los resultados confirman que rSFV permite la expresión transitoria de proteínas heterólogas en líneas celulares de hepatoma humano. Se necesitan estudios adicionales para determinar si la expresión disminuida de Core puede deberse a degradación de la proteína viral. Universidad Nacional de Colombia - Sede Bogotá - Facultad de Ciencias - Departamento de Biología 2020-09-09 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion application/pdf https://revistas.unal.edu.co/index.php/actabiol/article/view/79365 10.15446/abc.v26n1.79365 Acta Biológica Colombiana; Vol. 26 No. 1 (2021); 72 - 80 Acta Biológica Colombiana; Vol. 26 Núm. 1 (2021); 72 - 80 1900-1649 0120-548X spa https://revistas.unal.edu.co/index.php/actabiol/article/view/79365/pdf Derechos de autor 2020 Acta Biológica Colombiana https://creativecommons.org/licenses/by-nc-sa/4.0
institution UNAL
collection OJS
country Colombia
countrycode CO
component Revista
access En linea
databasecode rev-actabol
tag revista
region America del Sur
libraryname Sistema Nacional de Bibliotecas de la UNAL
language spa
format Digital
author Mira, Catalina
Yepes, Jesús Orlando
Henao, Luis Felipe
Montoya Guzmán, Melissa
Navas, Maria-Cristina
spellingShingle Mira, Catalina
Yepes, Jesús Orlando
Henao, Luis Felipe
Montoya Guzmán, Melissa
Navas, Maria-Cristina
Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus
author_facet Mira, Catalina
Yepes, Jesús Orlando
Henao, Luis Felipe
Montoya Guzmán, Melissa
Navas, Maria-Cristina
author_sort Mira, Catalina
title Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus
title_short Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus
title_full Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus
title_fullStr Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus
title_full_unstemmed Hepatitis C virus core protein expression in HepG2 cells using semliki forest virus
title_sort hepatitis c virus core protein expression in hepg2 cells using semliki forest virus
description The Hepatitis C Virus (HCV) encodes the structural protein Core, which in addition to being the capsid subunit, participates in different mechanisms of HCV infection pathogenesis. Since HCV in vitroreplication system has limitations, the use of viral vectors could be a useful tool to study the Core protein properties. To validate the Semliki Forest Virus (SFV) strategy in transduced HepG2 cells to study the HCV Core protein, the expression of green fluorescent protein (GFP) and Core protein expressions were detected 24 to 96 hours post-transduction in HepG2 cells transduced with rSFV. Core protein expression was lower than GFP expression in HepG2 cells. Since HCV Core protein can regulate the activity of the p53 gene, the transcriptional level of this gene was evaluated. A decrease in the level of p53 mRNA was observed in the cells after transduction, compared to the control cells. Although the cells transduced with rSFV-Core had the lowest level of p53 mRNA, the difference was not significant compared to cells transduced with rSFV-GFP. The results confirm that rSFV allows the transient expression of heterologous proteins in human hepatoma cell lines. Additional studies are needed to determine whether the decreased expression of Core may be due to the degradation of the viral protein. 
publisher Universidad Nacional de Colombia - Sede Bogotá - Facultad de Ciencias - Departamento de Biología
publishDate 2020
url https://revistas.unal.edu.co/index.php/actabiol/article/view/79365
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