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  • Original Paper
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Aberrant methylation of the ATM promoter correlates with increased radiosensitivity in a human colorectal tumor cell line

Oncogene volume 21, pages 3864–3871 (2002)Cite this article

Abstract

Recent findings suggest that DNA alkylating agents trigger cellular responses that overlap those activated after ionizing radiation. Moreover, activation of these responses is dependent upon a functional mismatch repair (MMR) system. These developments led us to test if MMR-deficient cells may be compromised in their ability to activate appropriate cellular signaling pathways after ionizing radiation. An initial experiment to address this notion was to determine the level of radiosensitivity of several MMR-deficient cell lines derived from patients with Hereditary Non-Polyposis Colorectal Cancer (HNPCC). While two of the three HNPCC lines investigated show levels of radiosensitivity consistent with that displayed by normal human fibroblasts, HCT-116 cells display moderate radiosensitivity compared to the other MMR-deficient lines. This increased sensitivity to ionizing radiation correlates with lowered levels of ATM expression in HCT-116. Analysis of genomic DNA from HCT-116 cells determined that these cells possess aberrant methylation of multiple CpG dinucleotides within the proximal promoter region of the ATM gene. The significance of this finding is underscored by our observations that co-culturing HCT-116 cells with the DNA demethylating agent 5-azacytidine reverses promoter methylation, promotes normal levels of ATM expression, and restores normal radiosensitivity. The proximal ATM promoter is a ∼520 bp region shared with the NPAT gene, and current evidence suggests that this region functions as a bi-directional promoter. We found that, unlike ATM, the methylation status of this intergenic region does not effect the expression of the NPAT gene. In sum, these observations indicate that the ATM gene is a novel target for epigentic silencing through inappropriate methylation of its proximal promoter region.

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Acknowledgements

We thank Dr Bryan Gebhardt (Department of Ophthamology, LSUHSC) for allowing us access to the 137Cs irradiator, Dr Timothy Jorgensen (Georgetown University) for advice in conducting radiosensitivity analyses, and Dr Wade Harper (Baylor College of Medicine) for his generous gift of anti-NPAT antisera. This work was supported by a research project grant (RPG) from the American Cancer Society (GMC-98564), and a grant from the Louisiana Board of Regents Health Excellence Fund.

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  1. Meena Shrivastav

    Present address: Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, Texas, USA

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, LSU Health Sciences Center, New Orleans, Louisiana, USA

    Wan-ju Kim, Quynh N Vo, Meena Shrivastav, Tamara A Lataxes & Kevin D Brown

  2. Stanley S Scott Cancer Center, LSU Health Sciences Center, New Orleans, Louisiana, USA

    Quynh N Vo, Meena Shrivastav, Tamara A Lataxes & Kevin D Brown

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  1. Wan-ju Kim
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Correspondence to Kevin D Brown.

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Kim, Wj., Vo, Q., Shrivastav, M. et al. Aberrant methylation of the ATM promoter correlates with increased radiosensitivity in a human colorectal tumor cell line. Oncogene 21, 3864–3871 (2002). https://doi.org/10.1038/sj.onc.1205485

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