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5-Fluorouracil signaling through a calcium–calmodulin-dependent pathway is required for p53 activation and apoptosis in colon carcinoma cells

Oncogene volume 32, pages 4529–4538 (2013)Cite this article

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Abstract

5-Fluorouracil (5-FU) is an anti-metabolite that is in clinical use for treatment of several cancers. In cells, it is converted into three distinct fluoro-based nucleotide analogs, which interfere with DNA synthesis and repair, leading to genome impairment and, eventually, apoptotic cell death. Current knowledge states that in certain cell types, 5-FU-induced stress is signaling through a p53-dependent induction of tumor necrosis factor-receptor oligomerization required for death-inducing signaling complex formation and caspase-8 activation. Here we establish a role of calcium (Ca2+) as a messenger for p53 activation in response to 5-FU. Using a combination of pharmacological and genetic approaches, we show that treatment of colon carcinoma cells stimulates entry of extracellular Ca2+ through long lasting-type plasma membrane channels, which further directs posttranslational phosphorylation of at least three p53 serine residues (S15, S33 and S37) by means of calmodulin (CaM) activity. Obstructing this pathway by the Ca2+-chelator BAPTA (1,2-bis(o-aminophenoxy)ethane- N,N,N',N'-tetraacetic acid) or by inhibitors of CaM efficiently reduces 5-FU-induced caspase activities and subsequent cell death. Moreover, ectopic expression of p53 S15A in HCT116 p53−/− cells confirmed the importance of a Ca2+–CaM–p53 axis in 5-FU-induced extrinsic apoptosis. The fact that a widely used therapeutic drug, such as 5-FU, is operating via this pathway could provide new therapeutic intervention points, or specify new combinatorial treatment regimes.

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Abbreviations

Ca2+:

calcium

DISC:

death-inducing signaling complex

5-FU:

5-fluorouracil.

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Acknowledgements

We thank Professors Bert Vogelstein, Peter H Krammer, Ladislav Anděra, Moshe Oren and Dr Inna Lavrik for their contribution of cells, antibodies and vectors. This study was supported by grants from the Swedish Science Foundation, the Swedish and Stockholm Cancer Societies, the Swedish Childhood Cancer Foundation and the EC-FP-6 (Chemores) and EC-FP-7 (APO-SYS). MO is a fellow of the Swedish Society of Medical Research.

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Author notes

  1. G Can and B Akpinar: These authors contributed equally to this work.

Authors and Affiliations

  1. Division of Toxicology, Institute of Environmental Medicine, Karolinska Institutet, Stockholm, Sweden

    G Can, B Akpinar, B Zhivotovsky & M Olsson

  2. Department of Molecular Biology and Genetics, Cancer Genetics and Molecular Hematology Laboratory, Izmir Institute of Technology, Izmir, Turkey

    Y Baran

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  1. G Can
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Correspondence to B Zhivotovsky.

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Can, G., Akpinar, B., Baran, Y. et al. 5-Fluorouracil signaling through a calcium–calmodulin-dependent pathway is required for p53 activation and apoptosis in colon carcinoma cells. Oncogene 32, 4529–4538 (2013). https://doi.org/10.1038/onc.2012.467

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