The biological effect of large single doses: a possible role for non-targeted effects in cell inactivation
Marlon R Veldwijk, Bo Zhang, Frederik Wenz, Carsten Herskind, Marlon R Veldwijk, Bo Zhang, Frederik Wenz, Carsten Herskind
Abstract
Background and purpose: Novel radiotherapy techniques increasingly use very large dose fractions. It has been argued that the biological effect of large dose fractions may differ from that of conventional fraction sizes. The purpose was to study the biological effect of large single doses.
Material and methods: Clonogenic cell survival of MCF7 and MDA-MB-231 cells was determined after direct X-ray irradiation, irradiation of feeder cells, or transfer of conditioned medium (CM). Cell-cycle distributions and the apoptotic sub-G1 fraction were measured by flow cytometry. Cytokines in CM were quantified by a cytokine antibody array. γH2AX foci were detected by immunofluorescence microscopy.
Results: The surviving fraction of MCF7 cells irradiated in vitro with 12 Gy showed an 8.5-fold decrease (95% c.i.: 4.4-16.3; P<0.0001) when the density of irradiated cells was increased from 10 to 50×10(3) cells per flask. Part of this effect was due to a dose-dependent transferrable factor as shown in CM experiments in the dose range 5-15 Gy. While no effect on apoptosis and cell cycle distribution was observed, and no differentially expressed cytokine could be identified, the transferable factor induced prolonged expression of γH2AX DNA repair foci at 1-12 h.
Conclusions: A dose-dependent non-targeted effect on clonogenic cell survival was found in the dose range 5-15 Gy. The dependence of SF on cell numbers at high doses would represent a "cohort effect" in vivo. These results support the hypothesis that non-targeted effects may contribute to the efficacy of very large dose fractions in radiotherapy.
Conflict of interest statement
Competing Interests: Dr. Zhang was supported by a fellowship from Shanghai Mai-Ge Medical Technology Co. (Shanghai, China). There are no patents, products in development or marketed products to declare. This does not alter the authors′ adherence to all the PLOS ONE policies on sharing data and materials.
Figures
References
- Hof H, Muenter M, Oetzel D, Hoess A, Debus J, et al. (2007) Stereotactic single-dose radiotherapy (radiosurgery) of early stage nonsmall-cell lung cancer (NSCLC). Cancer 110: 148–155.
- Kondziolka D, Flickinger JC, Lunsford LD (2012) Radiosurgery for brain metastases. Prog Neurol Surg 25: 115–122.
- Timmerman RD, Kavanagh BD, Cho LC, Papiez L, Xing L (2007) Stereotactic body radiation therapy in multiple organ sites. J Clin Oncol 25: 947–952.
- Morton G, Loblaw A, Cheung P, Szumacher E, Chahal M, et al. (2011) Is single fraction 15 Gy the preferred high dose-rate brachytherapy boost dose for prostate cancer? Radiother Oncol 100: 463–467.
- Vaidya JS, Joseph DJ, Tobias JS, Bulsara M, Wenz F, et al. (2010) Targeted intraoperative radiotherapy versus whole breast radiotherapy for breast cancer (TARGIT-A trial): an international, prospective, randomised, non-inferiority phase 3 trial. Lancet 376: 91–102.
- Veronesi U, Orecchia R, Luini A, Galimberti V, Zurrida S, et al. (2010) Intraoperative radiotherapy during breast conserving surgery: a study on 1,822 cases treated with electrons. Breast Cancer Res Treat 124: 141–151.
- Flickinger JC, Kondziolka D, Lunsford LD (2003) Radiobiological analysis of tissue responses following radiosurgery. Technol Cancer Res Treat 2: 87–92.
- Kirkpatrick JP, Meyer JJ, Marks LB (2008) The linear-quadratic model is inappropriate to model high dose per fraction effects in radiosurgery. Semin Radiat Oncol 18: 240–243.
- Herskind C, Wenz F (2009) Is there more to intraoperative radiotherapy (IORT) than physical dose ? Int J Radiat Oncol Biol 74: 976–977.
- Garcia LM, Wilkins DE, Raaphorst GP (2007) Alpha/beta ratio: A dose range dependence study. Int J Radiat Oncol Biol Phys 67: 587–593.
- Brenner DJ (2008) The linear-quadratic model is an appropriate methodology for determining isoeffective doses at large doses per fraction. Semin Radiat Oncol 18: 234–239.
- Buonanno M, de Toledo SM, Azzam EI (2011) Increased frequency of spontaneous neoplastic transformation in progeny of bystander cells from cultures exposed to densely ionizing radiation. PLoS One 6: e21540.
- Hei TK, Zhou H, Ivanov VN, Hong M, Lieberman HB, et al. (2008) Mechanism of radiation-induced bystander effects: a unifying model. J Pharm Pharmacol 60: 943–950.
- McMahon SJ, Butterworth KT, Trainor C, McGarry CK, O'Sullivan JM, et al. (2013) A kinetic-based model of radiation-induced intercellular signalling. PLoS One 8: e54526.
- Mothersill C, Seymour C (2012) Are epigenetic mechanisms involved in radiation-induced bystander effects? Front Genet 3: 74.
- Trainor C, Butterworth KT, McGarry CK, McMahon SJ, O'Sullivan JM, et al. (2012) DNA damage responses following exposure to modulated radiation fields. PLoS One 7: e43326.
- Blyth BJ, Sykes PJ (2011) Radiation-induced bystander effects: what are they, and how relevant are they to human radiation exposures? Radiat Res 176: 139–157.
- Gerashchenko BI, Howell RW (2003) Flow cytometry as a strategy to study radiation-induced bystander effects in co-culture systems. Cytometry A 54: 1–7.
- Gomez-Millan J, Katz IS, Farias Vde A, Linares-Fernandez JL, Lopez-Penalver J, et al. (2012) The importance of bystander effects in radiation therapy in melanoma skin-cancer cells and umbilical-cord stromal stem cells. Radiother Oncol 102: 450–458.
- Liu Z, Mothersill CE, McNeill FE, Lyng FM, Byun SH, et al. (2006) A dose threshold for a medium transfer bystander effect for a human skin cell line. Radiat Res 166: 19–23.
- Tomita M, Maeda M, Maezawa H, Usami N, Kobayashi K (2010) Bystander cell killing in normal human fibroblasts is induced by synchrotron X-ray microbeams. Radiat Res 173: 380–385.
- Yang H, Asaad N, Held KD (2005) Medium-mediated intercellular communication is involved in bystander responses of X-ray-irradiated normal human fibroblasts. Oncogene 24: 2096–2103.
- Liu Q, Schneider F, Ma L, Wenz F, Herskind C (2013) Relative Biologic Effectiveness (RBE) of 50 kV X-rays Measured in a Phantom for Intraoperative Tumor-Bed Irradiation. Int J Radiat Oncol Biol Phys 85: 1127–1133.
- Maier P, Fleckenstein K, Li L, Laufs S, Zeller WJ, et al. (2006) Overexpression of MDR1 using a retroviral vector differentially regulates genes involved in detoxification and apoptosis and confers radioprotection. Radiat Res 166: 463–473.
- Gow MD, Seymour CB, Ryan LA, Mothersill CE (2010) Induction of bystander response in human glioma cells using high-energy electrons: a role for TGF-beta1. Radiat Res 173: 769–778.
- Iyer R, Lehnert BE, Svensson R (2000) Factors underlying the cell growth-related bystander responses to alpha particles. Cancer Res 60: 1290–1298.
- Facoetti A, Mariotti L, Ballarini F, Bertolotti A, Nano R, et al. (2009) Experimental and theoretical analysis of cytokine release for the study of radiation-induced bystander effect. Int J Radiat Biol 85: 690–699.
- Mariotti LG, Bertolotti A, Ranza E, Babini G, Ottolenghi A (2012) Investigation of the mechanisms underpinning IL-6 cytokine release in bystander responses: The roles of radiation dose, radiation quality and specific ROS/RNS scavengers. Int J Radiat Biol 88: 751–762.
- Nagasawa H, Huo L, Little JB (2003) Increased bystander mutagenic effect in DNA double-strand break repair-deficient mammalian cells. Int J Radiat Biol 79: 35–41.
- Truman JP, Garcia-Barros M, Kaag M, Hambardzumyan D, Stancevic B, et al. (2010) Endothelial membrane remodeling is obligate for anti-angiogenic radiosensitization during tumor radiosurgery. PLoS One 5: e12310.
- Pomp J, Wike JL, Ouwerkerk IJ, Hoogstraten C, Davelaar J, et al. (1996) Cell density dependent plating efficiency affects outcome and interpretation of colony forming assays. Radiother Oncol 40: 121–125.
- Mothersill C, Seymour C (2001) Radiation-induced bystander effects: past history and future directions. Radiat Res 155: 759–767.
- Shareef MM, Cui N, Burikhanov R, Gupta S, Satishkumar S, et al. (2007) Role of tumor necrosis factor-alpha and TRAIL in high-dose radiation-induced bystander signaling in lung adenocarcinoma. Cancer Res 67: 11811–11820.
- Gow MD, Seymour CB, Byun SH, Mothersill CE (2008) Effect of dose rate on the radiation-induced bystander response. Phys Med Biol 53: 119–132.
- Shao C, Folkard M, Prise KM (2008) Role of TGF-beta1 and nitric oxide in the bystander response of irradiated glioma cells. Oncogene 27: 434–440.
- Zhou H, Ivanov VN, Lien YC, Davidson M, Hei TK (2008) Mitochondrial function and nuclear factor-kappaB-mediated signaling in radiation-induced bystander effects. Cancer Res 68: 2233–2240.
- Burdak-Rothkamm S, Short SC, Folkard M, Rothkamm K, Prise KM (2007) ATR-dependent radiation-induced gamma H2AX foci in bystander primary human astrocytes and glioma cells. Oncogene 26: 993–1002.
- Burdak-Rothkamm S, Rothkamm K, Prise KM (2008) ATM acts downstream of ATR in the DNA damage response signaling of bystander cells. Cancer Res 68: 7059–7065.
Source: PubMed