The influence of the time interval between the exposure to different quality radiation on murine B-16 melanoma cells survival

«Radiation and Risk», 2019, vol. 28, No. 1, pp.59-67

DOI: 10.21870/0131-3878-2019-28-1-59-67


Isaeva E.V. – Sen. Res., C. Sc., Vet.
Beketov E.E. – Head of Lab., C. Sc., Biol. Contacts: 4 Korolev str., Obninsk, Kaluga region, Russia, 249036. Tel.: +7-910-514-9947; e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it. .
Nasedkina N.V. – Lab. Ass.
Malakhov Y.P. – Res.
Troshina M.V. – Res.
Lychagin A.A. – Head of Lab., C. Sc., Phys.-Math.
Ulyanenko L.N. – Lead. Res., D. Sc., Biol., Prof.
Ulyanenko S.E. – Head of Dep., D. Sc., Biol.

A. Tsyb MRRC, Obninsk.


In modern radiotherapy schemes, radiations of different quality are used. Depending on the technical conditions, the irradiation sequence and interval between the exposures can be varied. For the combined gamma-neutron action, the dose contribution of neutrons varies from 20 to 40%. Taking into account the biophysical characteristics of charged particles and the processes of cellular damage and recovery, the dependence of biological effect on irradiation scheme is supposed. The influence of time interval between the irradiations of murine melanoma B-16 cells with 60-Co γ-rays with an average energy of 1.25 MeV and neutrons with an energy of 14.1 MeV was investigated. Portable neutron generator (NG-14) of continuous action, developed at the All-Russian Research Institute of Automation. N.L. Dukhova (Moscow) was a source of neutron radiation. Photons were provided by “Luch” installation. Irradiation was carried out in two sequences: first gamma radiation, then neutron (γ-n), and vice versa (n-γ); the interval between exposures was 30, 60 and 120 minutes. The effectiveness of irradiation was assessed by the cell survival according measured by clonogenic assay. It is shown that the biological effectiveness of neutrons and photons combined action did not depend on the sequence of irradiation. An increase in the time interval between irradiations (from 30 to 120 min) was accompanied with the growth of cell survival. Thus the time interval between promote the repair of non-lethal DNA damages. According to these results the most acceptable scheme of combined radiotherapy with neutrons and photons should include no more than 1 hour time interval between the irradiations with these agents.

Key words
Fast neutrons, NG-14, photons, combined action, irradiation sequence, synergism, DNA repair, clonogenic assay, melanoma, B-16 cells.


1. Wagner F.M., Specht H., Loeper-Kabasakal B., Breitkreutz H. Current state of therapy with fast neutrons. Sibirskiy onkologicheskiy zhurnal – Siberian Oncological Journal, 2015, no. 6, pp. 5-12. (In Russian).

2. Beketov E.E., Isaeva E.V., Koryakin S.N. Lychagin A.A., Ulyanenko S.E. Dependence of the efficiency of simultaneous action of gamma quanta and neutrons with energy of 14 MeV on the contribution of the densely ionizing component. Radiatsiya i risk – Radiation and Risk, 2012, vol. 21, no. 3, pp. 82-90. (In Russian).

3. Isaeva E.V., Beketov E.E., Koryakin S.N Lychagin A.A., Ulyanenko S.E. Comparison of biological efficiency of pulsed and continuous neutron radiation with energy of 14 MeV on the culture of cells of murine melanoma V-16. Radiatsiya i risk – Radiation and Risk, 2012, vol. 21, no. 2, pp. 83-90. (In Russian).

4. Lychagin A.A., Beketov E.E., Koryakin S.N., Isaeva E.V., Ulyanenko S.E. Experimental setup for irradiating biological objects with mixed fields of ionizing radiations of different quality. Meditsinskaya fizika – Medical Physics, 2013, vol. 59, no. 3, pp. 56-60. (In Russian).

5. Isaeva E.V., Beketov E.E., Koryakin S.N. Ulyanenko S.E., Lychagin A.A. A comparative study of the bio-logical effectiveness of 14-MEV neutron pulse and continuous radiation using mouse B-16 cells. Radiat. Prot. Dosimetry, 2014, vol. 161, no. 1-4, pp. 478-482.

6. Koryakin S.N., Kaydan N.A., Isaeva E.V., Ulyanenko L.N., Lychagin A.A., Ulyanenko S.E. Experience of using a portable domestic neutron generator in gamma-neutron therapy of domestic animals with malignant neoplasms. Radiatsiya i risk – Radiation and Risk, 2018, vol. 27, no. 1, pp. 94-106. (In Russian).

7. Mardynskiy Yu.S., Gulidov I.A., Aminov G.G., Ragulin Yu.A., Sysoev A.S. Combined (photon-neutron) therapy in complex treatment of locally advanced breast cancer. Voprosy onkologii – Oncology Questions, 2014, vol. 60, no. 4, pp. 489-492. (In  Russian).

8. Zaider M., Rossi H.H. The synergistic effect of different radiation. Radiat. Res., 1980, vol. 83, no. 3, pp. 732-739.

9. Beketov E.E. Effect of combined gamma-neutron irradiation on the death of tumor cells in vitro and in vivo. Cand. biol. sci. diss. Obninsk, 2012. 139 p. (In Russian).

10. Beketov E., Isaeva E., Malakhov E., Nasedkina N., Koryakin S., Ulyanenko S., Solovev A., Lychagin A. The study of biological effectiveness of U-70 accelerator carbon ions using melanoma B-16 clonogenic assay. Rad. Applic., 2017, vol. 2, no. 2, pp. 90-93.

11. McNally N., de Ronde J., Hinchliffe M. The effect of sequential irradiation with X-rays and fast neutrons on the survival of V79 Chinese hamster cells. Int. J. Radiat. Biol., 1984, vol. 45, no. 4, pp. 301-310.

12. Caswell R.S., Coyne J.J., Randolph M.Z. Kerma factors for neutron energies below 30 MeV. Radiat. Res., 1980, vol. 83, no. 2, pp. 217-254.

13. Staaf E., Brehwens K., Haghdoost S., Czub J., Wojcik A. Gamma-H2AX foci in cells exposed to a mixed beam of X-rays and alpha particles. Genome Integr., 2012, vol. 3, no. 1, pp. 8.

14. Sollazzo A., Shakeri-Manesh S., Fotouhi A., Czub J., Haghdoost S., Wojcik A. Interaction of low and high LET radiation in TK6 cells-mechanistic aspects and significance for radiation protection. J. Radiol. Prot., 2016, vol. 36, no. 4, pp. 721-735.

15. Cary L.H., Noutai D., Salber R.E., Williams M.S., Ngudiankama B.F., Whitnall M.H. Interactions between endothelial cells and T cells modulate responses to mixed neutron/gamma radiation. Radiat. Res., 2014, vol. 181, no. 6, pp. 592-604.

16. Zhang J., He Y., Shen X., Jiang D., Wang Q., Liu Q., Fang W. γ-H2AX responds to DNA damage induced by long-term exposure to combined low-dose-rate neutron and x-ray radiation. Mutat. Res. Genet. Toxicol. Environ. Mutagen., 2016, no. 795, pp. 36-40.

17. Ngo F., Blakely E., Tobias C. Sequential exposures of mammalian cells to low- and high-LET radiations. Radiat. Res., 1981, vol. 87, no. 1, pp. 59-78.

18. Joiner M., Brenner J., Denekamp J., Maughan E. The interaction between X-rays and 3 MeV neutrons in the skin of the mouse foot. Int. J. Radiat. Biol., 1984, vol. 46, no. 5, pp. 625-638.

19. Fesenko E.V., Luchnik N.V. Influence of combined gamma-neutron irradiation on the formation of structural mutations. Fundamental and applied aspects of neutron radiobiology. Collection of scientific works. Obninsk, SRIMR RAS USSR, 1985, pp. 54-64. (In Russian).

20. Tsyb T.S., Pakhomova O.N. Lethal effects of sequential action of gamma radiation and fast (0.85 MeV) reactor neutrons in yeast cells of Saccharomyces. 3rd Congress on Radiation Research. Pushchino, 1997, vol. 1, pp. 138-139. (In Russian).

21. Gajendiran N., Tanaka K., Kumaravel T., Kamada N. Neutron-induced adaptive response studied in go human lymphocytes using the comet assay. J. Radiat. Res., 2001, vol. 42, no. 1, pp. 91-101.

22. Vtyurin B.M., Ivanov V.K., Ivanov V.N., Medvedev V.S., Abdulkadirov S.A. Combined neutron and gamma-therapy of the cancer of the tongue. Meditsinskaya radiologiya – Medical Radiology, 1986, no. 9, pp. 14-19. (In Russian).

23. Higgins P., DeLuca P., Pearson D., Gould M. V79 survival following simultaneous or sequential irradiation by 15-MeV neutrons and 60Co photons. Radiat. Res., 1983, vol. 95, no. 1, pp. 45-56.

24. McNally N., de Ronde J., Hinchliffe M. Survival of V79 cells following simultaneous irradiation with X-rays and neutrons in air or hypoxia. Int. J. Radiat. Biol., 1985, vol. 48, no. 5, pp. 847-855.

25. Suzuki S. Survival of Chinese hamster V79 cells after irradiation with a mixture of neutrons and 60Co gamma-rays: experimental and theoretical analysis of mixed irradiation. Radiat. Res., 1993, vol. 133, no. 3, pp. 327-333.

26. Obaturov G.M., Sokolov V.A., Ul'yanenko S.Ye., Tsyb T.S. Actual problems of neutron radiobiology. Radiatsionnaya biologiya. Radioekologiya – Radiation Biology. Radioecology, 1997, vol. 37, no. 4, pp. 475-481.

27. Kaprin A.D., Galkin V.N., Zhavoronkov L.P., Ivanov V.K., Ivanov S.A., Romanko Yu.S. Synthesis of fundamental and applied research is the basis for ensuring a high level of scientific results and their introduction into medical practice. Radiatsiya i risk – Radiation and Risk, 2017, vol. 26, no. 2, pp. 26-40. (In Russian).

Full-text article (in Russian)