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

Acta Biológica Colombiana

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Title 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
 
Creator Mira, Catalina
Yepes, Jesús Orlando
Henao, Luis Felipe
Montoya Guzmán, Melissa
Navas, Maria-Cristina
 
Subject Transitory expression
p53
Viral Vector
HCV
Virology
Expresión transitoria
p53
Vector Viral
VHC
Virología
 
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. 
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.
 
Publisher Universidad Nacional de Colombia - Sede Bogotá - Faculdad de Ciencias - Departamento de Biología
 
Date 2020-09-09
 
Type info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
 
Format application/pdf
 
Identifier https://revistas.unal.edu.co/index.php/actabiol/article/view/79365
10.15446/abc.v26n1.79365
 
Source Acta Biológica Colombiana; Vol. 26 Núm. 1 (2021); 72 - 80
Acta Biológica Colombiana; Vol. 26 No. 1 (2021); 72 - 80
1900-1649
0120-548X
 
Language spa
 
Relation https://revistas.unal.edu.co/index.php/actabiol/article/view/79365/pdf
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Rights Derechos de autor 2020 Acta Biológica Colombiana
https://creativecommons.org/licenses/by-nc-sa/4.0
 

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