Περίληψη:
Background Hepatocellular carcinoma (HCC) is a cancer of poor prognosis,
with limited success in patient treatment, which it makes an excellent
target for gene therapy and viral oncolysis. Accordingly, herpes virus
simplex type-1 (HSV-1) is one of the most promising viral platforms for
transferring therapeutic genes and the development of oncolytic vectors
that can target, multiply in, and eradicate hepatoma cells via their
lytic cycle. Enhanced efficacy and specificity of HSV-1-based vectors
towards HCC may be achieved by using HCC-specific gene promoters to
drive selective viral gene expression and accomplish conditional
replication and/or to control the expression of therapeutic genes.
However, careful verification of promoter function in the context of the
replication-competent HSV-1 vectors is required. The present study aimed
to identify novel HCC-specific promoters that could efficiently direct
transgene expression to HCC cells and maintain their activity during
active viral replication.
Methods Publicly available microarray data from human HCC biopsies were
analysed in order to detect novel candidate genes induced primarily in
HCC compared to normal liver. HCC specificity and promoter activity were
evaluated by RT-PCR and chromatin immunoprecipitation. Additionally,
transcriptional activity of promoters was further evaluated in the
context of HSV-1 genome, using luciferase assays in cultured cells and
animal models.
Results Eight HCC-specific genes were characterised in this study:
Angiopoietin-like-3, Cytochrome P450, family 2, subfamily C, polypeptide
8, Vitronectin, Alcohol dehydrogenase 6-class V, Apolipoprotein B,
Fibrinogen beta chain, Inter-alpha-globulin-inhibitor H3 and
Inter-alpha-globulin-inhibitor H1. Specific HCC expression and active
gene transcription were confirmed in human liver and non-liver cell
lines and further evaluated in primary neoplastic cells from hepatitis C
and B virus (HCV-and HBV)-associated HCC patients. High promoter
activity and specificity in the presence of HSV-1 infection and from
within the viral genome, was validated, both in vitro and in vivo.
Conclusions We identified and experimentally characterized novel
hepatoma-specific promoters, which were valuable for cancer-specific
gene therapy, using HSV-1 vectors. Copyright (c) 2010 John Wiley &
Sons, Ltd.
Συγγραφείς:
Foka, Pelagia
Pourchet, Aldo
Hernandez-Alcoceba, Ruben and
Doumba, Polyxeni P.
Pissas, George
Kouvatsis, Vlassis and
Dalagiorgou, Georgia
Kazazi, Dorothea
Marconi, Peggy and
Foschini, Mariagiovanna
Manservigi, Roberto
Konstadoulakis,
Manousos M.
Koskinas, John
Epstein, Alberto L.
Mavromara,
Penelope
