Abstract
Metastatic melanoma is a deadly cancer that fails to respond to conventional chemotherapy and is poorly understood at the molecular level1. p53 mutations often occur in aggressive and chemoresistant cancers but are rarely observed in melanoma1,2. Here we show that metastatic melanomas often lose Apaf-1, a cell-death effector that acts with cytochrome c and caspase-9 to mediate p53-dependent apoptosis3. Loss of Apaf-1 expression is accompanied by allelic loss in metastatic melanomas, but can be recovered in melanoma cell lines by treatment with the methylation inhibitor 5-aza-2′-deoxycytidine (5aza2dC). Apaf-1-negative melanomas are invariably chemoresistant and are unable to execute a typical apoptotic programme in response to p53 activation. Restoring physiological levels of Apaf-1 through gene transfer or 5aza2dC treatment markedly enhances chemosensitivity and rescues the apoptotic defects associated with Apaf-1 loss. We conclude that Apaf-1 is inactivated in metastatic melanomas, which leads to defects in the execution of apoptotic cell death. Apaf-1 loss may contribute to the low frequency of p53 mutations observed in this highly chemoresistant tumour type.
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Acknowledgements
We thank B. Stillman and N. K. Cheung for support; M. McCurrach, W. Jiang, L. Chen and M. Dudas for technical assistance; M. Zhang, R. Davuluri and I. Ioshikhes for analysis of Apaf-1 CpG islands and assembly of Apaf-1 genomic sequence; G. Núñez for Apaf-1XL pcDNA construct; E. Querido, P. E. Branton and F. L. Graham for Ad-p53 and Ad-LacZ; and A. Houghton and R. Camalier for melanoma cell lines. We also thank G. Hannon, C. Schmitt, C. Hazan, K. Pohar, T. Tiongson and the Cold Spring Harbor Genome Center for help and advice. Y.L. is a Pew Scholar and S.W.L. is a Rita Allen Scholar. This work was supported by a long-term Postdoctoral Fellowship from the Human Frontiers in Science Program and a Special Fellowship from the Leukemia and Lymphoma Society (M.S.S), a K08 grant (D.P.), an ASCO Young Investigator Award (J.M.), grants from the Seraph Foundation (YL), and from the NIH and the NCI (C.C.C. and S.W.L.).
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Soengas, M., Capodieci, P., Polsky, D. et al. Inactivation of the apoptosis effector Apaf-1 in malignant melanoma . Nature 409, 207–211 (2001). https://doi.org/10.1038/35051606
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DOI: https://doi.org/10.1038/35051606
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