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  • Original Article
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Inhibition of Siah ubiquitin ligase function

Abstract

Tumor hypoxia induces the upregulation of hypoxia-inducible factor 1α (Hif-1α), which in turn induces the expression of genes including VEGF to recruit new blood vessel outgrowth, enabling tumor growth and metastasis. Interference with the Hif-1 pathway and neoangiogenesis is an attractive antitumor target. The hydroxylation of Hif-1α by prolyl-hydroxylase (PHD) proteins during normoxia serves as a recognition motif for its proteasomal degradation. However, under hypoxic conditions, hydroxylation is inhibited and furthermore, PHD proteins are themselves polyubiquitylated and degraded by Siah ubiquitin ligases. Our data demonstrate for the first time that inhibition of the interaction between Siah and PHD proteins using a fragment derived from a Drosophila protein (phyllopod) interferes with the PHD degradation. Furthermore, cells stably expressing the phyllopod fragment display reduced upregulation of Hif-1α protein levels and Hif-1-mediated gene expression under hypoxia. In a syngeneic mouse model of breast cancer, the phyllopod fragment reduced tumor growth and neoangiogenesis and prolonged survival of the mice. In addition, levels of Hif-1α and its target Glut-1 are reduced in tumors expressing the phyllopod fragment. These data show, in a proof-of-principle study, that Siah protein, the most upstream component of the hypoxia pathway yet identified, is a viable drug target for antitumor therapies.

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Acknowledgements

The EO771 cells were a kind gift from Dr Robin Anderson, Peter MacCallum Cancer Centre. The expression plasmid for FLAG-TIEG-1 was provided by Steven Johnsen and Thomas Spelsburg (Mayo Clinic and Foundation, Rochester, NY, USA). The expression plasmid for Myc-FIH-1 was a kind gift from Daniel Peet (School of Molecular and Biomedical Science, University of Adelaide, Adelaide, Australia). This study was funded by grants of the Australian National Health and Medical Research Council (NHMRC) to AM, CH and DB (no. 400321 and no. 509331).

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Correspondence to D D L Bowtell.

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Möller, A., House, C., Wong, C. et al. Inhibition of Siah ubiquitin ligase function. Oncogene 28, 289–296 (2009). https://doi.org/10.1038/onc.2008.382

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