INVESTIGADORES
CUESTAS Maria Lujan
congresos y reuniones científicas
Título:
Transient expression of the NS5a protein of Hepatitis C virus (HCV) in HeLa cells
Autor/es:
MONTENEGRO G, FERNÁNDEZ GAMBA A, RIVERO C, MINASSIAN M, RUIZ V, CUESTAS M, TRINKS J, MATHET V, OUBIÑA J
Lugar:
Pinamar, Argentina
Reunión:
Congreso; XLI Reunión Anual de SAIB; 2005
Institución organizadora:
Sociedad Argentina de Investigación Bioquímica y Biología Molecular
Resumen:
INTRODUCTION: It is uncertain whether the HCV NS5A protein up- or down-regulates the expression of anti-apoptotic proteins, as well as if plays any role in the cellular immortalization event. OBJECTIVE: To investigate the effect of transiently transfected HCV NS5A protein in the expression of cell cycle regulators. MATERIAL AND METHODS: pDest12.2-NS5A recombinant vector was transfected in HeLa cells using Lipofectamine 2000. Appropriate controls were used. Cells were studied 24 hours after transfection. The expression of NS5A, c-Myc, cIAP1, cIAP2, Bcl-XL/XS, Tankyrase (up-regulator of telomerase) and Gankyrin was measured by Western blot. Primary specific mono- and polyclonal antibodies as well as secondary polyclonal antibodies were used throughout, and revealed by chemiluminescense using the highly sensitive ECL Plus system. RESULTS: The expression of NS5A was detected in transiently transfected cells. No differences were recorded between such cells and controls regarding the expression of cellular proteins involved in cell cycle regulation or immortalization. CONCLUSIONS: HeLa cells transiently transfected with HCV NS5A protein do not show detectable changes in the level of expression of the studied proteins. Apoptotic stimuli are being investigated to further evaluate the participation of this protein in apoptosis regulation using a stable inducible expression system.