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Medium-mediated intercellular communication is involved in bystander responses of X-ray-irradiated normal human fibroblasts

Oncogene volume 24, pages 2096–2103 (2005)Cite this article

Abstract

Although radiation-induced bystander effects have been demonstrated in a number of cell types, the studies have largely been performed using high linear energy transfer (LET) radiation, such as α-particles. The literature is contradictory on whether fibroblasts show bystander responses, especially after low LET radiation such as X- or γ-rays and whether the same signal transmission pathways are involved. Herein, a novel transwell insert culture dish method is used to show that X-irradiation induces medium-mediated bystander effects in AGO1522 normal human fibroblasts. The frequency of micronuclei formation in unirradiated bystander cells increases from a background of about 6.5% to about 9–13% at all doses from 0.1 to 10 Gy to the irradiated cells. Induction of p21Waf1 protein and foci of γ-H2AX in bystander cells is also independent of dose to the irradiated cells above 0.1 Gy. In addition, levels of reactive oxygen species (ROS) were increased persistently in directly irradiated cells up to 60 h after irradiation and in bystander cells for 30 h. Adding Cu–Zn superoxide dismutase (SOD) and catalase to the medium decreases the formation of micronuclei and induction of p21Waf1 and γ-H2AX foci in bystander cells, suggesting oxidative metabolism plays a role in the signaling pathways in bystander cells. The results of clonogenic assay of bystander cells showed that survival of bystander cells decreases from 0 to 0.5 Gy, and then is independent of the dose to irradiated cells from 0.5 to 10 Gy. Unlike the response with p21Waf1 expression, γ-H2AX foci and micronuclei, adding SOD and catalase has no effect on the survival of bystander cells. The data suggest that irradiated cells release toxic factors other than ROS into the medium.

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Acknowledgements

We thank Drs Aloke Chatterjee, Howard Liber and Betsy Sutherland for helpful discussions. This research was supported in part by the Office of Science (BER), US Department of Energy Grant No. DE-FG02-02ER63304 and NASA Grant No NAG 2-1642 (to KDH).

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  1. Nesrin Asaad

    Present address: Department of Radiology, UMDNJ-New Jersey Medical School, Newark, NJ, 07103, USA

Authors and Affiliations

  1. Department of Radiation Oncology, Harvard Medical School, Massachusetts General Hospital, COX 302, Fruit Street, Boston, MA, 02114, USA

    Hongying Yang, Nesrin Asaad & Kathryn D Held

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  1. Hongying Yang
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Correspondence to Kathryn D Held.

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Yang, H., Asaad, N. & Held, K. Medium-mediated intercellular communication is involved in bystander responses of X-ray-irradiated normal human fibroblasts. Oncogene 24, 2096–2103 (2005). https://doi.org/10.1038/sj.onc.1208439

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