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Implication of functional activity for determining therapeutic efficacy of suicide genes in vitro

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Abstract

Gene-Directed Enzyme Prodrug Therapy (GDEPT), commonly known as suicide gene therapy, provides a selective approach to eradicate tumor cells that is currently considered as an alternate approach to conventional therapy for cancers due to its high efficacy. Herein, we have demonstrated functional activity of the cytosine deaminase (CD) and the hybrid cytosine deaminase-uracil phosphoribosyltransferase (CD-UPRT) suicide genes in transfected cell lines. We have monitored retention profiles of various metabolites that were formed during enzymatic conversion of the prodrug 5-flurocytosine (5-FC) using reverse phase HPLC method. Therapeutic effect of suicide genes was established by cell viability and toxicity assay, whereas apoptotic induction was confirmed by DNA fragmentation ELISA. Our results demonstrated that 5-FC/CD-UPRT-mediated apoptotic cell death was more than 5-FC/CD, which could be further potentiated with anticancer compound curcumin. Such results corroborated 5-FC/CD-UPRT in combination with curcumin as a better chemosensitization method.

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Acknowledgements

This work was financially supported by the Department of Biotechnology [BT/PR9988/NNT/28/76/2007] and Council of Scientific and Industrial Research [No.37 (1248)/06/EMRII] Government of India.

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Authors and Affiliations

  1. Department of Biotechnology, Indian Institute of Technology Guwahati, Guwahati, 781039, India

    P. Gopinath & Siddhartha Sankar Ghosh

  2. Centre for Nanotechnology, Indian Institute of Technology Guwahati, Guwahati, 781039, India

    Siddhartha Sankar Ghosh

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  1. P. Gopinath
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  2. Siddhartha Sankar Ghosh
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Correspondence to Siddhartha Sankar Ghosh.

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Gopinath, P., Ghosh, S.S. Implication of functional activity for determining therapeutic efficacy of suicide genes in vitro. Biotechnol Lett 30, 1913–1921 (2008). https://doi.org/10.1007/s10529-008-9787-1

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  • DOI: https://doi.org/10.1007/s10529-008-9787-1

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