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Increased sensitivity of glioma cells to 5-fluorocytosine following photo-chemical internalization enhanced nonviral transfection of the cytosine deaminase suicide gene

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Abstract

Despite advances in surgery, chemotherapy and radiotherapy, the outcomes of patients with GBM have not significantly improved. Tumor recurrence in the resection margins occurs in more than 80 % of cases indicating aggressive treatment modalities, such as gene therapy are warranted. We have examined photochemical internalization (PCI) as a method for the non-viral transfection of the cytosine deaminase (CD) suicide gene into glioma cells. The CD gene encodes an enzyme that can convert the nontoxic antifungal agent, 5-fluorocytosine, into the chemotherapeutic drug, 5-fluorouracil. Multicell tumor spheroids derived from established rat and human glioma cell lines were used as in vitro tumor models. Plasmids containing either the CD gene alone or together with the uracil phosphoribosyl transferase (UPRT) gene combined with the gene carrier protamine sulfate were employed in all experiments.PCI was performed with the photosensitizer AlPcS2a and 670 nm laser irradiance. Protamine sulfate/CD DNA polyplexes proved nontoxic but inefficient transfection agents due to endosomal entrapment. In contrast, PCI mediated CD gene transfection resulted in a significant inhibition of spheroid growth in the presence of, but not in the absence of, 5-FC. Repetitive PCI induced transfection was more efficient at low CD plasmid concentration than single treatment. The results clearly indicate that AlPcS2a-mediated PCI can be used to enhance transfection of a tumor suicide gene such as CD, in malignant glioma cells and cells transfected with both the CD and UPRT genes had a pronounced bystander effect.

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Acknowledgments

This work was supported by grants from the Norwegian Radium Hospital Research Foundation and the Chao Cancer Center. Portions of this work were made possible through access to the Laser Microbeam and Medical Program (LAMMP) and the Chao Cancer Center Optical Biology Shared Resource at the University of California, Irvine.

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Authors and Affiliations

  1. Beckman Laser Institute and Medical Clinic, University of California Irvine, 1002 Health Sciences Rd, Irvine, CA, 92617, USA

    Frederick Wang, Genesis Zamora, Chung-Ho Sun, Anthony Trinidad & Henry Hirschberg

  2. Department of Chemical Engineering/Material Science, University of California Irvine, Irvine, CA, USA

    Changho Chun & Young Jik Kwon

  3. Department of Pharmaceutical Sciences, University of California Irvine, Irvine, CA, USA

    Young Jik Kwon

  4. Department of Radiation Biology, The Norwegian Radium Hospital, Oslo University Hospital, Oslo, Norway

    Kristian Berg

  5. Department of Health Physics and Diagnostic Sciences, University of Nevada, Las Vegas, NV, USA

    Steen J. Madsen & Henry Hirschberg

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  1. Frederick Wang
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Correspondence to Frederick Wang.

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Frederick Wang and Genesis Zamora have contributed equally to this Study.

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Wang, F., Zamora, G., Sun, CH. et al. Increased sensitivity of glioma cells to 5-fluorocytosine following photo-chemical internalization enhanced nonviral transfection of the cytosine deaminase suicide gene. J Neurooncol 118, 29–37 (2014). https://doi.org/10.1007/s11060-014-1410-9

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  • DOI: https://doi.org/10.1007/s11060-014-1410-9

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