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Enhancement of radiosensitivity by 5-Aza-CdR through activation of G2/M checkpoint response and apoptosis in osteosarcoma cells

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Tumor Biology

Abstract

Radiation resistance is a major problem preventing successful treatment. Therefore, identifying sensitizers is vitally important for radiotherapy success. Epigenetic events such as DNA methylation have been proposed to mediate the sensitivity of tumor therapy. In this study, we investigated the influence of demethylating agent 5-Aza-2′-deoxycytidine (5-Aza-CdR) on the radiosensitivity of human osteosarcoma cell lines. 5-Aza-CdR was capable of sensitizing three osteosarcoma cells to irradiation in a time-dependent manner, with the maximum effect attained by 48 h. Pretreatment with 5-Aza-CdR synchronized cells in G2/M phase of the cell cycle and enhanced irradiation-induced apoptosis compared with irradiation alone in SaOS2, HOS, and U2OS cells. Moreover, 5-Aza-CdR restored mRNA expressions of 14-3-3σ, CHK2, and DAPK-1 in the three cells, accompanied with demethylation of their promoters. These findings demonstrate that demethylation with 5-Aza-CdR increases radiosensitivity in some osteosarcoma cells through arresting cells at G2/M phase and increasing apoptosis, which is partly mediated by upregulation of 14-3-3σ, CHK2, and DAPK-1 genes, suggesting that 5-Aza-CdR may be a potential radiosensitizer to improve the therapy effect in osteosarcoma.

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Abbreviations

5-Aza-CdR:

5-Aza-2′-deoxycytidine

DNMT:

DNA methyltransferase

IC20 :

Concentration to cause a 20 % reduction in cell proliferation

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

D0 :

Dose to reduce survival to 37 %

SF2:

Surviving fraction at 2 Gy

SER:

Sensitizer enhancement ratio

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Acknowledgments

This study was funded by the National Natural Science Foundation of China (81201756) and Natural Science Foundation Project of YN CSTC (2011FZ316 and 2012FD090).

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Authors and Affiliations

  1. Key Laboratory of Oncology, Cancer Center, Division of Internal Medicine, Chinese PLA General Hospital and Chinese PLA Medical School, Beijing, 100853, China

    Yi Li & PeiLiang Geng

  2. Department of Oncology, Kunming General Hospital of Chengdu Military Command, Kunming, 650032, China

    Yi Li, Xu Lin, Jun Liu, Lichun Huang, Bin Su & Hong Chen

  3. Department of Orthopedics, The Affiliated Hospital of Academy of Military Medical Sciences, PLA 307th Hospital, Beijing, 100071, China

    Weihao Jiang

  4. Department of Radiation Oncology, Chinese PLA general Hospital, Beijing, 100853, China

    Yunlai Wang

  5. Department of Oncology, PLA No. 161 Center Hospital, Wuhan, 430012, China

    Jie Yao

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  1. Yi Li
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  2. PeiLiang Geng
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  3. Weihao Jiang
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  10. Hong Chen
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Corresponding author

Correspondence to Hong Chen.

Additional information

Yi Li and PeiLiang Geng contributed equally to this work.

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Li, Y., Geng, P., Jiang, W. et al. Enhancement of radiosensitivity by 5-Aza-CdR through activation of G2/M checkpoint response and apoptosis in osteosarcoma cells. Tumor Biol. 35, 4831–4839 (2014). https://doi.org/10.1007/s13277-014-1634-5

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  • DOI: https://doi.org/10.1007/s13277-014-1634-5

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