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RUNX3 inhibits hypoxia-inducible factor-1α protein stability by interacting with prolyl hydroxylases in gastric cancer cells

Oncogene volume 33, pages 1458–1467 (2014)Cite this article

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Abstract

RUNX3 is silenced by histone modification and hypoxia-inducible factor (HIF)-1α is stabilized under hypoxia, but little is known of cross-talk between RUNX3 and HIF-1α under hypoxia. In the present study, the authors investigated the effect of RUNX3 on HIF-1α stability in gastric cancer cells. RUNX3 overexpression was found to downregulate HIF-1α stability under normoxic and hypoxic conditions. Furthermore, the activity of a luciferase reporter containing five copies of vascular endothelial growth factor (VEGF) promoter hypoxia-responsive element (5 × HRE) and the amount of secreted VEGF, were diminished in RUNX3-expressing but increased in RUNX3-knockdown cells. When expression of RUNX3 was recovered using epigenetic reagents the expressions of HIF-1α and VEGF were clearly suppressed under hypoxic conditions. RUNX3 also significantly attenuated the half-life of HIF-1α protein, and induced the cytosolic localization and ubiquitination of HIF-1α. In addition, RUNX3 directly interacted with the C-terminal activation domain of HIF-1α and prolyl hydroxylase (PHD) 2 and enhanced the interaction between HIF-1α and PHD2, which potentiated proline hydroxylation and promoted the degradation of HIF-1α. Furthermore, RUNX3 overexpression significantly inhibited hypoxia-induced angiogenesis in vitro and in vivo. Taken together, these results suggest that RUNX3 destabilizes HIF-1α protein by promoting the proline hydroxylation of HIF-1α through binding to HIF-1α/PHD2. RUNX3 appears to be a novel suppressor of HIF-1α and of hypoxia-mediated angiogenesis in gastric cancer cells.

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Acknowledgements

This work was supported by a National Research Foundation grant from the Basic Research Lab program (2012-028835) and from Mid-career Researcher Program (2010-0026741) funded by the Korean Ministry of Education, Science Technology (MEST).

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Authors and Affiliations

  1. School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University, Daegu, Republic of Korea

    S H Lee

  2. School of Medicine, Chungbook National University, Cheongju, Republic of Korea

    S C Bae

  3. College of Pharmacy, Seoul National University, Seoul, Republic of Korea

    K W Kim

  4. College of Pharmacy, Kyungpook National University, Daegu, Republic of Korea

    S H Lee & Y M Lee

  5. Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, Republic of Korea

    S H Lee & Y M Lee

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Lee, S., Bae, S., Kim, K. et al. RUNX3 inhibits hypoxia-inducible factor-1α protein stability by interacting with prolyl hydroxylases in gastric cancer cells. Oncogene 33, 1458–1467 (2014). https://doi.org/10.1038/onc.2013.76

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