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Plasmid injection and application of electric pulses alter endogenous mRNA and protein expression in B16.F10 mouse melanomas

Cancer Gene Therapy volume 17, pages 864–871 (2010)Cite this article

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Abstract

The application of electric pulses to tissues causes cell membrane destabilization, allowing exogenous molecules to enter the cells. This delivery technique can be used for plasmid gene therapy. Reporter gene expression after plasmid delivery with eight representative published protocols was compared in B16.F10 mouse melanoma tumors. This expression varied significantly based on the pulse parameters utilized for delivery. To observe the possible influence of plasmid injection and/or pulse application on endogenous gene expression, levels of stress-related mRNAs 4 and 24 h after delivery were determined by PCR array. Increases in mRNA levels for several inflammatory chemokines and cytokines were observed in response to plasmid injection, electric pulses alone or the combination. This upregulation was confirmed by individual real-time reverse transcription TaqMan PCR assays. Proteins were extracted at the same time points from identically treated tumors and inflammatory protein levels were assayed by enzyme-linked immunosorbent assay and by a custom multiplex bead array. Increases in inflammatory protein levels generally paralleled mRNA levels. Some differences were observed, which may have been due to differing expression kinetics. The observed upregulated expression of these cytokines and chemokines may aid or inhibit the therapeutic effectiveness of immune-based cancer gene therapies.

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Acknowledgements

The work in this study was supported by NIH 1 R21 CA106860 (LH).

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Authors and Affiliations

  1. Department of Medical Laboratory and Radiation Sciences, Frank Reidy Research Center for Bioelectrics, Old Dominion University, Norfolk, VA, USA

    L C Heller & R Heller

  2. Department of Molecular Medicine, University of South Florida College of Medicine, Tampa, FL, USA

    Y L Cruz, B Ferraro & H Yang

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  1. L C Heller
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  2. Y L Cruz
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  3. B Ferraro
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  5. R Heller
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Correspondence to L C Heller.

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RH has ownership interest in RMR Technologies and owns stock and stock options of Inovio Biomedical Corporation.

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Heller, L., Cruz, Y., Ferraro, B. et al. Plasmid injection and application of electric pulses alter endogenous mRNA and protein expression in B16.F10 mouse melanomas. Cancer Gene Ther 17, 864–871 (2010). https://doi.org/10.1038/cgt.2010.43

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