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Low-dose metronomic cyclophosphamide treatment mediates ischemia-dependent K-ras mutation in colorectal carcinoma xenografts

Oncogene volume 27, pages 3729–3738 (2008)Cite this article

Abstract

Antiangiogenic therapies are promising approaches to cancer control, but the details of their effects on subsequent tumor progression are not fully understood. Such therapies have the potential to eventually generate extensive amounts of tumor ischemia, and we previously demonstrated that ischemic conditions induce K-ras mutations in cells with deficient mismatch repair (MMR) mechanisms. This suggested that similar effects on oncogene mutagenesis may accompany antiangiogenic therapy. To test this, MMR-deficient colorectal cancer cells (Dks-8) were xenografted into immune-deficient mice and treated with the antiangiogenic regimen of low-dose/metronomic cyclophosphamide for 2 weeks followed by a 2-week recovery period without therapy. This treatment resulted in transient tumor growth inhibition, increased hypoxia, and decreased microvessel density, and cancer cells from treated tumors acquired activating mutations of the K-ras oncogene (K-rasG13D). In vitro exposure of Dks-8 cells to the active metabolite of cyclophosphamide (4-hydroxycyclophosphamide) had no effect on the K-ras status, indicating that there was no direct action of this alkylating agent on K-ras mutagenesis. In addition, cells sorted from hypoxic regions of Dks-8 tumors were enriched in K-rasG13D mutants. Collectively, our studies suggest that increases in tumor hypoxia induced by antiangiogenic treatment may lead to K-ras mutation and consequently tumor progression, especially in susceptible individuals.

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Acknowledgements

We thank Courtney Stone and Kanwal Minhas for technical support, Barb Mitchell for mouse husbandry and the members of the Coomber laboratory for their valuable suggestions. This work was supported by Canadian Cancer Society (National Cancer Institute of Canada Grant no. 016117 to BL Coomber and JW Rak).

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Authors and Affiliations

  1. Department of Biomedical Sciences, University of Guelph, Guelph, Ontario, Canada

    S Shahrzad & B L Coomber

  2. Department of Cell Biology, School of Medicine, Fukuoka University, Fukuoka, Japan

    S Shirasawa

  3. Research Institute, International Medical Center of Japan, Tokyo, Japan

    T Sasazuki

  4. Department of Pediatrics, Montreal Children's Hospital, McGill University, Montreal, Quebec, Canada

    J W Rak

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Correspondence to B L Coomber.

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Supplementary Information accompanies the paper on the Oncogene website (http://www.nature.com/onc).

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Shahrzad, S., Shirasawa, S., Sasazuki, T. et al. Low-dose metronomic cyclophosphamide treatment mediates ischemia-dependent K-ras mutation in colorectal carcinoma xenografts. Oncogene 27, 3729–3738 (2008). https://doi.org/10.1038/sj.onc.1211031

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