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MDA-7/IL-24, a novel tumor suppressor/cytokine is ubiquitinated and regulated by the ubiquitin–proteasome system, and inhibition of MDA-7/IL-24 degradation enhances the antitumor activity

Cancer Gene Therapy volume 15, pages 1–8 (2008)Cite this article

Abstract

Steady-state protein levels are determined by the balance between protein synthesis and degradation. Protein half-lives are determined primarily by degradation, and the major degradation pathways involve either lysosomal destruction or an ATP-dependent process involving ubiquitination to target proteins to the proteosome. Studies have shown that multiple tumor-suppressor proteins are ubiquitinated and degraded by the 26S proteasome. In the present study, we investigated whether the tumor suppressor/cytokine melanoma differentiation-associated gene-7/interleukin-24 gene (MDA-7/IL-24) protein is ubiquitinated and its degradation controlled by the proteasome. Treatment of ovarian (2008) and lung (H1299) tumor cells with adenoviral delivery of mda-7 (Ad-mda7) or Ad-mda7 plus the proteosome inhibitor MG132 showed that MDA-7 protein expression was dependent upon proteosome activity. Western blot and immunoprecipitation analyses verified that the MDA-7 protein was ubiquitinated and that ubiquitinated-MDA-7 levels were increased in MG132-treated cells. These results were confirmed using small interfering RNA (siRNA)-mediated knockdown of ubiquitin. Furthermore, ubiquitinated MDA-7 protein was degraded by the 26S proteasome, as MDA-7 accumulation was observed only when cells were treated with MG132 but not with lysosome or protease inhibitors. Inhibition of the catalytic β-5 subunit of the 20S proteasome using siRNA resulted in MDA-7 protein accumulation. Finally, treatment of tumor cells with Ad-mda7 plus the proteasome inhibitor bortezomib resulted in increased tumor cell killing. Our results show that MDA-7/IL-24 is ubiquitinated and degraded by the 26S proteasome. Furthermore, inhibition of MDA-7 degradation results in enhanced tumor killing, identifying a novel anticancer strategy.

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Acknowledgements

We thank Debbie Smith for manuscript preparation, Michael Worley and the Department of Scientific Publications at MD Anderson Cancer Center for editorial assistance. This study was supported by funds from the National Cancer Institute grants RO1-CA 102716 and PO1 CA 06294 (R Ramesh) and CA 89778, CA 88421 and CA 097598 (S Chada); Cancer Center Support grant CA 16672 and a sponsored research agreement with Introgen Therapeutics, Inc. We like to disclose financial interest; RR is a consultant for Introgen Therapeutics and SC is an employee of Introgen Therapeutics, Inc.

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  1. B Gopalan

    Present address: 4Present address: Institute of Bioengineering and Nanotechnology, Biopolis Way, Singapore 138669.,

  2. B Gopalan and M Shanker: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Thoracic and Cardiovascular Surgery, The University of Texas, MD Anderson Cancer Center, Houston, TX, USA

    B Gopalan, M Shanker, A Scott, C D Branch & R Ramesh

  2. Introgen Therapeutics, Inc., Houston, TX, USA

    S Chada

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Correspondence to R Ramesh.

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Gopalan, B., Shanker, M., Scott, A. et al. MDA-7/IL-24, a novel tumor suppressor/cytokine is ubiquitinated and regulated by the ubiquitin–proteasome system, and inhibition of MDA-7/IL-24 degradation enhances the antitumor activity. Cancer Gene Ther 15, 1–8 (2008). https://doi.org/10.1038/sj.cgt.7701095

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