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Synergistic growth inhibition based on small-molecule p53 activation as treatment for intraocular melanoma

Oncogene volume 31, pages 1105–1116 (2012)Cite this article

Abstract

The prognosis of patients with uveal melanoma is poor. Because of the limited efficacy of current treatments, new therapeutic strategies need to be developed. Because p53 mutations are uncommon in uveal melanoma, reactivation of p53 may be used to achieve tumor regression. We investigated the use of combination therapies for intraocular melanoma, based on the p53 activators Nutlin-3 and reactivation of p53 and induction of tumor cell apoptosis (RITA) and the topoisomerase I inhibitor Topotecan. Nutlin-3 treatment induced p53-dependent growth inhibition in human uveal melanoma cell lines. The sensitivity to Nutlin-3 of the investigated cell lines did not correlate with basal Hdm2 or Hdmx levels. Nutlin-3 synergized with RITA and Topotecan to induce apoptosis in uveal melanoma cell lines and short-term cultures. Drug synergy correlated with enhanced induction of p53–Ser46 phosphorylation, which was attenuated by ATM inhibition. Nutlin-3 and Topotecan also significantly delayed tumor growth in vivo in a murine B16F10 model for ocular melanoma. Combination treatment appeared to inhibit tumor growth slightly more efficient than either drug alone. Nutlin-3, RITA and Topotecan lead to comparable p53 activation and growth inhibition under normoxia and hypoxia. Treatment with Nutlin-3 or RITA had no effect on HIF-1α induction by hypoxia, whereas the combination of these two drugs did inhibit hypoxia-induced HIF-1α. Also Topotecan, alone or in combination with Nutlin-3, reduced HIF-1α protein levels, suggesting that a certain level of DNA damage response is required for p53-mediated downregulation of HIF-1α. In conclusion, combination treatments based on small-molecule-induced p53 activation may have clinical potential for uveal melanoma.

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Acknowledgements

We thank Dr PA van der Velden and M Versluis for the primer sets to detect the expression of the HIF target genes and the early cultures of primary uveal melanoma cells, Dr A Vertegaal for the antibodies detecting HIF-1α and HIF-2α proteins, Prof BR Ksander for providing the Mel cell lines, Dr G Selivanova for providing RITA and Dr A Levine for providing anti-Mdm2 antibody. This study was supported by grants from the Netherlands Organization for Scientific Research NWO (Mozaiek Grant 017.003.059) and by EC FP6 funding (contract 503576).

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Authors and Affiliations

  1. Department of Molecular Cell Biology, Leiden University Medical Center, Leiden, The Netherlands

    J de Lange, K Lodder, M Verlaan-de Vries, A F A S Teunisse & A G Jochemsen

  2. Department of Ophthalmology, Leiden University Medical Center, Leiden, The Netherlands

    L V Ly & M J Jager

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  1. J de Lange
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Correspondence to A G Jochemsen.

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de Lange, J., Ly, L., Lodder, K. et al. Synergistic growth inhibition based on small-molecule p53 activation as treatment for intraocular melanoma. Oncogene 31, 1105–1116 (2012). https://doi.org/10.1038/onc.2011.309

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