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Adenovirus‐mediated cytosine deaminase/5‐fluorocytosine suicide gene therapy of human hepatoblastoma in vitro
Author(s) -
Warmann Steven W.,
Armeanu Sorin,
Heigoldt Heike,
Ruck Peter,
Vonthein Reinhard,
Heitmann Heike,
Seitz Guido,
Lemken MarieLuise,
Bitzer Michael,
Fuchs Jörg,
Lauer Ulrich M.
Publication year - 2009
Publication title -
pediatric blood and cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.116
H-Index - 105
eISSN - 1545-5017
pISSN - 1545-5009
DOI - 10.1002/pbc.21956
Subject(s) - cytosine deaminase , suicide gene , hepatoblastoma , genetic enhancement , transduction (biophysics) , cancer research , prodrug , cell culture , medicine , cytosine , microbiology and biotechnology , gene , biology , pharmacology , genetics , biochemistry
Background Multidrug resistance is a key factor for the sobering outcome of relapsed and metastatic human hepatoblastoma (HB). Gene directed treatment approaches were recently identified as possible treatment options against advanced HB, in which standard chemotherapy regimens are partially insufficient. The aim of this study was to systematically analyze the effects of suicide gene therapy in three HB cell lines using a yeast‐derived cytosine deaminase (YCD)‐combined yeast uracil phosphoribosyltransferase (YUPRT)‐based adenovirus‐mediated gene transfer. Procedure YCD and YUPRT were fused to form the bifunctional suicide gene SuperCD. Adeonoviral vectors were used for transduction. Tumor cells transduced at MOI 50 were incubated with 5‐fluorocytosine (5‐FC) in ascending concentrations. Results Transduction rates were 87.8% (+6.7) in the mixed HB cell line HUH6, 98.6% (+1.4) in the epithelial HB cell line HepT1 and 93.6% (+0.6) in the multifocal HB embryonal cell line HepT3, respectively. In HepT3 and HepT1 cells suicide gene therapy with SuperCD/5‐FC was highly effective leading to HB cell damage far above those of application of the prodrug 5‐FC only. In HUH6 cells the approach had no effect due to a lack in activity of the CMV promoter being employed for transcription of the SuperCD transgene. Conclusion Assuming employment of fully active promoters, the SuperCD/5‐FC approach may serve as a potentially useful anti‐tumor strategy against advanced HB. Pediatr Blood Cancer 2009;53:145–151. © 2009 Wiley‐Liss, Inc.