Recombinant adenoviral vector containing tumor-specific L-plastin promoter fused to cytosine deaminase gene as a transcription unit: generation and functional test.
The expression of therapeutic transgenes in recombinant adenoviral vectors is a major cause of toxicity in dividing cancer cells as well as non dividing normal cells. To solve the problem of toxicity to normal cells, we have reported on a recombinant adenoviral vector system (AdLP-) in which the expression of the transgene is directed by the tumor-specific L-plastin promoter (LP) (Chung et al., 1999). The object of this study was to generate a recombinant adenoviral vector system which would generate tumor cell specific expression of cytosine deaminase (CD) gene. We report the construction of a replication-incompetent adenoviral vector in which CD is driven by the L-plastin promoter (AdLPCD). Infection of 293 cells by AdLPCD generated the functional CD protein as measured by HPLC analysis for the conversion of 5-Fluorocytosine (5-FC) to 5-Fluorouracil (5-FU). HPLC analysis in conjunction with counting radioactivity for [6-3H]-5FC and [6-3H]-5FU demonstrated vector dose-dependent conversion of 5-FC to 5-FU in AdLPCD infected ovarian cancer cells. The results from present and previous studies (Peng et al., 2001; Akbulut et al., 2003) suggest that the use of the AdLPCD/5-FC system may be of value in the treatment of cancer including microscopic ovarian cancer in the peritoneal cavity.
College of Pharmacy, Duksung Women's University, Seoul, Korea.
Injae
I
Chung
Albert B
AB
Deisseroth
D000256:Adenoviridae / Q000235:genetics*; D019677:Artificial Gene Fusion; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid; D043525:Cytosine Deaminase / Q000096:biosynthesis / Q000235:genetics*; D005260:Female; D005437:Flucytosine / Q000493:pharmacokinetics; D005472:Fluorouracil / Q000378:metabolism; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D006801:Humans; D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D010051:Ovarian Neoplasms; D010750:Phosphoproteins / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic
D008562:Membrane Glycoproteins; D008840:Microfilament Proteins; D010750:Phosphoproteins; C026742:plastin; D005437:Flucytosine; D043525:Cytosine Deaminase; D005472:Fluorouracil