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Transforming growth factor-β gene silencing using adenovirus expressing TGF-β1 or TGF-β2 shRNA

Cancer Gene Therapy volume 20, pages 94–100 (2013)Cite this article

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Abstract

Tumor cells secrete a variety of cytokines to outgrow and evade host immune surveillance. In this context, transforming growth factor-β1 (TGF-β1) is an extremely interesting cytokine because it has biphasic effects in cancer cells and normal cells. TGF-β1 acts as a growth inhibitor in normal cells, whereas it promotes tumor growth and progression in tumor cells. Overexpression of TGF-β1 in tumor cells also provides additional oncogenic activities by circumventing the host immune surveillance. Therefore, this study ultimately aimed to test the hypothesis that suppression of TGF-β1 in tumor cells by RNA interference can have antitumorigenic effects. However, we demonstrated here that the interrelation between TGF-β isotypes should be carefully considered for the antitumor effect in addition to the selection of target sequences with highest efficacy. The target sequences were proven to be highly specific and effective for suppressing both TGF-β1 mRNA and protein expression in cells after infection with an adenovirus expressing TGF-β1 short hairpin RNA (shRNA). A single base pair change in the shRNA sequence completely abrogated the suppressive effect on TGF-β1. Surprisingly, the suppression of TGF-β1 induced TGF-β3 upregulation, and the suppression of TGF-β2 induced another unexpected downregulation of both TGF-β1 and TGF-β3. Taken together, this information may prove useful when considering the design for a novel cancer immunogene therapy.

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Abbreviations

TGF-β:

transforming growth factor-β

shRNA:

short hairpin RNA

GFP:

green fluorescence protein

ELISA:

enzyme-linked immunosorbent assay

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Acknowledgements

This work was supported by the Industrial Strategic Technology Development program (10035562: Development of nucleic acid-based anticancer drugs overcoming the immunotherapy resistance) funded by the Ministry of Knowledge Economy (MKE, Korea). This work was also supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) and funded by the Ministry of Education. Science and Technology (2012-0002108). S Oh and E Kim are funded by the Brain Korea 21 project for Medical Science, Yonsei University, College of Medicine, Seoul, South Korea.

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Authors and Affiliations

  1. Institute for Cancer Research, College of Medicine, Yonsei University, Seoul, South Korea

    S Oh, E Kim, D Kang, M Kim & J J Song

  2. Brain Korea 21 Project for Medical Science, College of Medicine, Yonsei University, Seoul, South Korea

    S Oh & E Kim

  3. Department of Oncology, Affiliated Hospital of Yanbian University, Yanji, Jilin Province, People’s Republic of China

    D Kang

  4. Department of Internal Medicine, College of Medicine, Yonsei University, Seoul, South Korea

    J-H Kim

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Correspondence to J-H Kim or J J Song.

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Oh, S., Kim, E., Kang, D. et al. Transforming growth factor-β gene silencing using adenovirus expressing TGF-β1 or TGF-β2 shRNA. Cancer Gene Ther 20, 94–100 (2013). https://doi.org/10.1038/cgt.2012.90

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