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Antibody-directed double suicide gene therapy targeting of MUC1- positive leukemia cells in vitro and in vivo
- Source :
- Current gene therapy. 13(5)
- Publication Year :
- 2013
-
Abstract
- Our aim was to specifically transfer the cytosine deaminase (CD) and thymidine kinase (TK) genes into mucin 1 (MUC1)-positive leukemia cells by anti-MUC1 antibody directed infection of replication-defective lentivirus and to evaluate the targeted cytotoxicity of double suicide genes to leukemia. The target gene vector (containing CD and TK) and envelope (containing GFP and anti-MUC1) and packaging plasmids were cotransfected into 293T cells to produce the recombinant lentivirus. Suicide genes in virus-infected leukemia cells (U937, Jurkat, and K562) were detected by western blot. The cytotoxicity and bystander effect in vitro and the therapeutic effect in vivo were detected after treatment with the prodrugs. The results revealed that combined treatment with prodrug 5-fluorocytosine (5-FC) and ganciclovir (GCV) inhibited leukemia cell growth and caused significant bystander effect than treatment with either prodrug alone. TK/GCV treatment alone induced degeneration and cell death while the effect of CD/5-FC alone mainly caused vacuolar degeneration and necrosis. The addictive effects of combinatorial use of GCV and 5-FC mainly induced swelling of the mitochondria followed by necrosis of the leukemia cells. In vivo experiments revealed that both single and combinatorial prodrug treatments could prolong the survival time of leukemic mice. In summary, anti-MUC1 antibody directed lentiviral vector successfully transduced dual suicide genes and exerted targeted cytotoxicity against MUC1 positive leukemia cells. This targeted lentiviral dual suicide gene delivering system provides a promising approach for clinical treatment of leukemia in future.
- Subjects :
- Male
Genetic Vectors
Flucytosine
Mice, Nude
Apoptosis
Biology
Jurkat cells
Thymidine Kinase
Viral vector
Cytosine Deaminase
Jurkat Cells
Mice
Drug Discovery
Genetics
medicine
Animals
Humans
Prodrugs
Cytotoxicity
Molecular Biology
Ganciclovir
Genetics (clinical)
Cell Proliferation
Leukemia
Cytosine deaminase
Lentivirus
Mucin-1
Gene Transfer Techniques
Genes, Transgenic, Suicide
Genetic Therapy
U937 Cells
Suicide gene
medicine.disease
Molecular biology
Disease Models, Animal
HEK293 Cells
Thymidine kinase
Gene Targeting
Molecular Medicine
K562 Cells
K562 cells
Plasmids
Subjects
Details
- ISSN :
- 18755631
- Volume :
- 13
- Issue :
- 5
- Database :
- OpenAIRE
- Journal :
- Current gene therapy
- Accession number :
- edsair.doi.dedup.....213138f4fcf4e81eebd03a1657c1f3ba