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Constitutive nitric oxide acting as a possible intercellular signaling molecule in the initiation of radiation-induced DNA double strand breaks in non-irradiated bystander cells

Oncogene volume 26, pages 2330–2339 (2007)Cite this article

Abstract

The initiation and propagation of the early processes of bystander signaling induced by low-dose α-particle irradiation are very important for understanding the underlying mechanism of the bystander process. Our previous investigation showed that the medium collected from cell culture exposed to low-dose α-particle rapidly induced phosphorylated form of H2AX protein foci formation among the non-irradiated medium receptor cells in a time-dependent manner. Using NG-methyl-L-arginine, 4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate and Nω-nitro-L-arginine (L-NNA) treatment before exposure to 1 cGy α-particle, we showed in the present study that nitric oxide (NO•) produced in the irradiated cells was important and necessary for the DNA double strand break inducing activity (DIA) of conditioned medium and the generation of NO• in irradiated confluent AG1522 cells is in a time-dependent manner and that almost all NO• was generated within 15 min post-irradiation. Concurrently, the kinetics of NO• production in the medium of irradiated cells after irradiation was rapid and in a time-dependent manner as well, with a maximum yield observed at 10 min after irradiation with electron spin resonance analysis. Furthermore, our results that 7-Nitroindazole and L-NNA, but not aminoguanidine hemisulfate, treatment before exposure to 1 cGy α-particle significantly decrease the DIA of the conditioned medium suggested that constitutive NO• from the irradiated cells possibly acted as an intercellular signaling molecule to initiate and activate the early process (⩽30 min) of bystander response after low-dose irradiation.

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Abbreviations

AG:

aminoguanidine hemisulfate

c-PTIO:

2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide

DAF-FM diacetate:

4-amino-5-methylamino-2′,7′-difluorofluorescein diacetate

DIA:

DSB-inducing activity (of conditioned medium collected from the irradiated culture)

D-NMMA:

NG-methyl-D-arginine

DSBs:

DNA double strand breaks

ESR:

electron spin resonance

γ-H2AX:

phosphorylated form of H2AX protein

L-NMMA:

NG-methyl-L-arginine

L-NNA:

Nω-nitro-L-arginine

7-Ni:

7-Nitroindazole

NO•:

nitric oxide

NO2−:

nitrite

NOS:

nitric oxide synthase

ONOO−:

peroxynitrite

RIBE:

radiation-induced bystander effects

RNS:

reactive nitrogen species

ROS:

reactive oxygen species

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Acknowledgements

We thank Drs Mohammad Athar, Hari Bhat and Vladimir Ivanov for critical reading of the paper and their helpful discussions, and Dr Haiying Hang for his technical assistance with DSB detection. This work was funded by National Nature Science Foundation of China under Grant nos. 10225526 and 30570435, Grant KSCX2-SW-324 of Chinese Academy of Sciences, and US National Institutes of Health Grants ES 012888 and Environmental Center Grant ES09089.

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Authors and Affiliations

  1. Key Laboratory of Ion Beam Bioengineering, Institute of Plasma Physics, Chinese Academy of Sciences, Hefei, People's Republic of China

    W Han, L Wu, S Chen, L Bao, L Zhang, E Jiang, Y Zhao, A Xu & Z Yu

  2. Center for Radiological Research, College of Physicians and Surgeons,Columbia University, New York, NY, USA

    T K Hei

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Correspondence to L Wu.

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

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Han, W., Wu, L., Chen, S. et al. Constitutive nitric oxide acting as a possible intercellular signaling molecule in the initiation of radiation-induced DNA double strand breaks in non-irradiated bystander cells. Oncogene 26, 2330–2339 (2007). https://doi.org/10.1038/sj.onc.1210024

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