Journal
MOLECULAR AND CELLULAR BIOCHEMISTRY
Volume 324, Issue 1-2, Pages 21-29Publisher
SPRINGER
DOI: 10.1007/s11010-008-9980-5
Keywords
Suicide gene therapy; Apoptosis; CD-UPRT; Bystander effects; Atomic force microscope
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Funding
- Department of Biotechnology [BT/PR9988/NNT/28/76/2007]
- Council of Scientific and Industrial Research [37 ( 1248)/06/EMR-II]
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Cytosine deaminase-uracil phosphoribosyl transferase (CD-UPRT) fusion gene is known to exhibit therapeutic effect by inducing apoptosis in vitro. However, bystander effects of 5-flurocytosine (5-FC)/CD-UPRT and the molecular mechanism for apoptosis are yet to be established. In the present study, we have generated BHK21 cell line expressing both CD-UPRT and green fluorescent protein (GFP) from two separate transcripts, where GFP was used as a noninvasive probe to monitor the therapeutic effect of CD-UPRT. Enzyme activity of CD-UPRT in the stable cell line was measured by the reverse phase high-performance liquid chromatography analysis. Inhibition of cell growth and strong bystander effects of 5-FC/CD-UPRT were established, whereas characteristic surface morphology of apoptotic cell death was identified by AFM analysis. Involvement of various apoptotic signaling genes using semi-quantitative RT-PCR has been explored to substantiate the potential application of 5-FC/CD-UPRT suicide gene in therapy.
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