In vitro gene targeting in human hepatoblastoma

Poor treatment results in advanced hepatoblastoma (HB) made alternative treatment approaches desirable. Gene-directed tumor therapy is increasingly investigated in different malignancies. The aim of this study was to analyze possible alternatives of gene transfer into HB cells and to study therapeut...

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Bibliographic Details
Published in:Pediatric surgery international 2006-01, Vol.22 (1), p.16-23
Main Authors: Warmann, Steven W, Armeanu, Sorin, Frank, Heike, Buck, Heike, Graepler, Florian, Lemken, Marie-Luise, Heitmann, Heike, Seitz, Guido, Lauer, Ulrich M, Bitzer, Michael, Fuchs, Jörg
Format: Article
Language:English
Subjects:
Adaptor Proteins, Signal Transducing - genetics
Adenoviridae
Cell Line, Tumor
Gene Expression Regulation, Neoplastic
Gene Silencing
Gene Targeting - methods
Gene Transfer Techniques
Genes, Transgenic, Suicide
Genetic Vectors
Hepatoblastoma - genetics
Hepatoblastoma - therapy
Humans
Liposomes - therapeutic use
Liver Neoplasms - genetics
Liver Neoplasms - therapy
Organ Specificity
Promoter Regions, Genetic
Sendai virus
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Summary:Poor treatment results in advanced hepatoblastoma (HB) made alternative treatment approaches desirable. Gene-directed tumor therapy is increasingly investigated in different malignancies. The aim of this study was to analyze possible alternatives of gene transfer into HB cells and to study therapeutic applications based on different strategies. Liposomal transfection of HB cells was assessed using liver-specific promoters, and adenovirus and Sendai virus transductions were performed in vitro. Transfer efficiencies were measured via flow cytometry determining expression of vector-encoded marker gene green fluorescent protein. Gene silencing of the anti-apoptotic bcl-2 gene in HUH6 cells was performed using lipofection of small interfering RNA (siRNA). Additionally, suicide gene therapy was carried out through a yeast-derived cytosine deaminase (YCD)-combined yeast uracil phosphoribosyltransferase (YUPRT)-based adenovirus-mediated gene transfer, leading to a potent intracellular prodrug transformation of 5-fluorocytosine into 5-fluorouracil. Treatment efficiencies were monitored via MTT viability assay. Highest gene transfer rates (86%) were observed using adenovirus transduction. We furthermore observed a significant therapeutic effect of adenovirus-mediated YCD::YUPRT suicide gene transfer. Liposomal-mediated anti-bcl-2 siRNA transfer led to a significant improvement of cisplatin treatment in HUH6 cells. Liver-specific promoters were found to be strongly active in HUH6 cells (mixed HB-derived), but less active in HepT1 cells (embryonal HB-derived). Liposomal transfection and viral transduction are effective approaches to genetically manipulate HB cells in vitro. For the first time, we demonstrate a positive effect of siRNA gene silencing in this malignancy. Additionally, we successfully investigated a model of adenovirus-based suicide gene therapy in HB cell cultures. Our data strongly encourage further studies assessing these alternative treatment approaches.
ISSN:0179-0358
1437-9813
DOI:10.1007/s00383-005-1573-8