HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct

  • Authors:
    • Outi Meriläinen
    • Tanja Hakkarainen
    • Tiina Wahlfors
    • Riikka Pellinen
    • Jarmo Wahlfors
  • View Affiliations

  • Published online on: July 1, 2005     https://doi.org/10.3892/ijo.27.1.203
  • Pages: 203-208
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Abstract

Protein transduction domain (PTD) from HIV-1 TAT protein has been reported to translocate across the mammalian cell membrane, also as a part of fusion proteins. However, the true nature of TAT-mediated intercellular spreading is still under debate because it has been claimed to be a fixation artifact. To study the spreading of TAT fusion proteins and their potency to enhance thymidine kinase/ ganciclovir (HSV-TK/GCV) cancer gene therapy, we constructed a novel triple fusion protein containing TAT PTD, HSV-TK and green fluorescent protein (TAT-TK-GFP). This fusion protein has three functional domains in the same polypeptide, allowing reliable determination of the relationship between transduction rate and cell killing efficiency. TAT-TK-GFP was cloned into a lentivirus vector and used for analyses of TAT-mediated protein translocation and enhancement of HSV-TK/GCV cytotoxicity. The triple fusion protein was expressed correctly in vitro, but cell-to-cell translocation was not observed in rat glioma cells (BT4C). However, TAT-TK-GFP made BT4C and SKOV3.ip1 (human ovarian carcinoma) cells significantly more sensitive to ganciclovir than TK-GFP, whereas the effect in PC-3 human prostate carcinoma cells was more subtle. It was also observed that growth in lower serum concentration (2.5-5%) abolished the enhancement in BT4C cells, suggesting that high proliferation rate is one of the factors that contribute to TAT PTD-mediated enhancement of cytotoxicity. In summary, our results indicate that TAT PTD fusion proteins do not translocate intercellularly at detectable levels, but enhancement of the HSV-TK/GCV cytotoxicity can be detected in rat and human tumor cell lines in vitro.

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July 2005
Volume 27 Issue 1

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Meriläinen O, Hakkarainen T, Wahlfors T, Pellinen R and Wahlfors J: HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct. Int J Oncol 27: 203-208, 2005.
APA
Meriläinen, O., Hakkarainen, T., Wahlfors, T., Pellinen, R., & Wahlfors, J. (2005). HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct. International Journal of Oncology, 27, 203-208. https://doi.org/10.3892/ijo.27.1.203
MLA
Meriläinen, O., Hakkarainen, T., Wahlfors, T., Pellinen, R., Wahlfors, J."HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct". International Journal of Oncology 27.1 (2005): 203-208.
Chicago
Meriläinen, O., Hakkarainen, T., Wahlfors, T., Pellinen, R., Wahlfors, J."HIV-1 TAT protein transduction domain mediates enhancement of enzyme prodrug cancer gene therapy in vitro: a study with TAT-TK-GFP triple fusion construct". International Journal of Oncology 27, no. 1 (2005): 203-208. https://doi.org/10.3892/ijo.27.1.203