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Malignant melanoma cells acquire resistance to DNA interstrand cross-linking chemotherapeutics by p53-triggered upregulation of DDB2/XPC-mediated DNA repair

Oncogene volume 33, pages 1964–1974 (2014)Cite this article

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Abstract

Malignant melanoma is a cancer characterized by high chemoresistance although p53 is rarely mutated. Here, we show that p53 wild-type melanoma cells acquire resistance to cell death induced by fotemustine (FM), which is a representative of alkylating DNA interstrand cross-linking agents used in melanoma therapy. We show that drug-induced resistance is a result of p53-dependent upregulation of the nucleotide excision repair (NER) genes xeroderma pigmentosum complementation group C (XPC) and damaged DNA-binding protein 2 (DDB2), which stimulate the repair of DNA interstrand cross-links (ICLs) arising from O6-chloroethylguanine. Consequently, TP53 mutated cells are unable to repair ICLs, leading to prolonged ATM, ATR and checkpoint kinase 1 (CHK1) activation, and finally apoptosis. The roles of p53 and NER in ICL-triggered cell death were confirmed by knockdown of p53 and XPC. Upregulation of XPC and DDB2 in p53wt cells following a single drug treatment is a robust and sustained response that lasts for up to 1 week. Pretreatment with an inducing dose followed by a high and toxic dose of FM provoked an adaptive response as the killing outcome of the challenge dose was reduced. Upregulation of XPC and DDB2 was also observed in a melanoma mouse xenograft model following systemic administration of FM. Additionally, XPC and DDB2 induction occurred upon treatment with other cross-linking anticancer drugs, such as cisplatin and mafosfamide, indicating it is a general response of cancer cells to this group of chemotherapeutics. Collectively, the data indicate that p53-dependent upregulation of XPC and DDB2 is a key mechanism upon genotoxic stress, whereby melanoma cells acquire resistance towards DNA cross-linking agents. To our knowledge, this is the first demonstration of upregulation of NER following a single dose of a DNA interstrand cross-linker, which is a robust and long-lasting effect that impacts the killing response of cancer cells to subsequent treatments.

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Acknowledgements

We gratefully acknowledge a gift of DDB2 antibodies from Dr Vesna Rapic-Otrin, University of Pittsburgh and Dr Beate Köberle for her help. We thank Rebekka Kitzinger for technical assistance, Georg Nagel for his help with animal experiments and Christian Henninger for helpful advice concerning tumor sample processing. Work was supported by DFG KA724 and RO3617.

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  1. C Barckhausen and W P Roos: These authors contributed equally to this work

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  1. Institute of Toxicology, Medical Center of the University Mainz, Obere Zahlbacher Str. 67, Mainz, Germany

    C Barckhausen, W P Roos, S C Naumann & B Kaina

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  1. C Barckhausen
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  2. W P Roos
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Correspondence to B Kaina.

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Barckhausen, C., Roos, W., Naumann, S. et al. Malignant melanoma cells acquire resistance to DNA interstrand cross-linking chemotherapeutics by p53-triggered upregulation of DDB2/XPC-mediated DNA repair. Oncogene 33, 1964–1974 (2014). https://doi.org/10.1038/onc.2013.141

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