Acquired resistance of B16 tumor cells to protons after prolonged fractional electron irradiation

«Radiation and Risk», 2020, vol. 29, No. 4, pp.69-83

DOI: 10.21870/0131-3878-2020-29-4-69-83

Authors

Beketov E.E. – Head of Lab., C. Sc., Biol. Contacts: 4 Korolev str., Obninsk, Kaluga region, Russia, 249035. Tel.: +7910-514-9947; e-mail: This email address is being protected from spambots. You need JavaScript enabled to view it. .
Isaeva E.V. – Sen. Res., C. Sc., Vet.
Nasedkina N.V. – Jun. Res.
Zamulaeva I.A. – Head of Dep., D. Sc., Biol., Prof.
Matchuk O.N. – Sen. Res., C. Sc., Biol.
Ulyanenko L.N. – Lead. Res., D. Sc., Biol., Prof.
Malakhov E.P. – Jun. Res.
Kisel A.A. – Jun. Res.
Golovanova O.Yu. – Engineer-physicist
Ulyanenko S.E. – Head of Dep., D. Sc., Biol.
Ivanov S.A. – Director, MD, Prof. A. Tsyb MRRC.
Shegai P.V. – Head of the Center for Innovative Radiological and Regenerative Technologies, C. Sc., Med.
Kaprin A.D. – General Director, Academician of RAS, MD, Prof. NMRRC.

¹ A. Tsyb MRRC, Obninsk
² NMRRC Russian Ministry of Health, Moscow

Abstract

Cancer radiotherapy effectiveness largely depends on tumor cells radiosensitivity. Inherent or acquired radioresistance of tumor cells is important challenge in radiation therapy. Response of tumor cells to fractionated radiation therapy has been investigated by many research groups. At present time the use of protons for cancer research and treatment has expanded rapidly. In this connection research on sensitivity of tumor cells to proton beam therapy is an urgent task. The aim of the study was to assess sensitivity of irradiated with electrons or protons B16 melanoma cells to the next electron beam or proton beam irradiation at comparable total doses. Studies with the use of stable tumor cell lines with acquired radioresistance may be useful for the development of effective treatment plan tailored to the patients with relapses or metastases that have occurred after prior unsuccessful radiotherapy with standard types of radiation. Protons were provided by Prometeus installation scanning beam and the electron beam of the accelerator Novac-11. Cells radiosensitivity was measured by clonogenic assay. The resistance of cells first irradiated with protons and electrons to the next irradiation with protons and electrons was estimated by clonogenic assay. DNA damages, cell size, proliferative activity and cell cycle phase distribution were also evaluated. The study demonstrated that fractionated irradiation of B16 cells with electrons at the total dose of 60 Gy causes significant reduction of cells radiosensitivity to the next irradiation with protons, radiosensitivity of irradiated cells to the second irradiation with electrons remains the same. In contrast, the first fractionated irradiation of cells with protons at the total dose of 50 Gy does not affect the radiosensitivity of the cells to the next irradiation with electrons or protons.

Key words
protons, electrons, radiation therapy, fractionation, radioresistance, B16 melanoma cells, comet assay, clonogenic activity.

References

1. Kuwahara Y., Roudkenar M.H., Urushihara Y., Saito Y., Tomita K., Roushandeh A.M., Sato T., Kurimasa A., Fukumoto M. Clinically relevant radioresistant cell line: a simple model to understand cancer radiore-sistance. Med. Mol. Morphol., 2017, vol. 50, no. 4, pp. 195-204.

2. Fukuda K., Sakakura C., Miyagawa K., Kuriu Y., Kin S., Nakase Y., Hagiwara A., Mitsufuji S., Okazaki Y., Hayashizaki Y., Yamagishi H. Differential gene expression profiles of radioresistant oesophageal cancer cell lines established by continuous fractionated irradiation. Br. J. Cancer, 2004, vol. 91, no. 8, pp. 1543-1550.

3. Jing Z., Gong L., Xie C.Y., Zhang L., Su H.F., Deng X., Wu S.X. Reverse resistance to radiation in KYSE-150R esophageal carcinoma cell after epidermal growth factor receptor signal pathway inhibition by ce-tuximab. Radiother. Oncol., 2009, vol. 93, no. 3, pp. 468-473.

4. Kuwahara Y., Li L., Baba T., Nakagawa H., Shimura T., Yamamoto Y., Ohkubo Y., Fukumoto M. Clinically relevant radioresistant cells efficiently repair DNA double-strand breaks induced by X-rays. Cancer Sci., 2009, vol. 100, no. 4, pp. 747-752.

5. Kuwahara Y., Mori M., Oikawa T., Shimura T., Ohtake Y., Mori S., Ohkubo Y., Fukumoto M. The modified high-density survival assay is the useful tool to predict the effectiveness of fractionated radiation exposure. J. Radiat. Res., 2010, vol. 51, no. 3, pp. 297-302.

6. Lynam-Lennon N., Reynolds J.V., Pidgeon G.P., Lysaght J., Marignol L., Maher S.G. Alterations in DNA repair efficiency are involved in the radioresistance of esophageal adenocarcinoma. Radiat. Res., 2010, vol. 174, no. 6, pp. 703-711.

7. Qing Y., Yang X.Q., Zhong Z.Y., Lei X., Xie J.Y., Li M.X., Xiang D.B., Li Z.P., Yang Z.Z., Wang G., Wang D. Microarray analysis of DNA damage repair gene expression profiles in cervical cancer cells radioresistant to 252Cf neutron and X-rays. BMC Cancer, 2010, vol. 10, pp. 71.

8. Russell J., Wheldon T.E., Stanton P. A radioresistant variant derived from a human neuroblastoma cell line is less prone to radiation-induced apoptosis. Cancer Res., 1995, vol. 55, no. 21, pp. 4915-4921.

9. Sato K., Imai T., Okayasu R., Shimokawa T. Heterochromatin domain number correlates with X-ray and carbonion radiation resistance in cancer cells. Radiat. Res., 2014, vol. 182, no. 4, pp. 408-419.

10. Wei K., Kodym R., Jin C. Radioresistant cell strain of human fibrosarcoma cells obtained after long-term exposure to x-rays. Radiat. Environ. Biophys., 1998, vol. 37, no. 2, pp. 133-137.

11. Xie L., Song X., Yu J., Wei L., Song B., Wang X., Lv L. Fractionated irradiation induced radio-resistant esophageal cancer EC109 cells seem to be more sensitive to chemotherapeutic drugs. J. Exp. Clin. Cancer Res., 2009, vol. 28, no. 1, pp. 68.

12. Pearce A.G., Segura T.M., Rintala A.C., Rintala-Maki N.D., Lee H. The generation and characterization of a radiation-resistant model system to study radioresistance in human breast cancer cells. Radiat. Res., 2001, vol. 156, no. 6, pp. 739-750.

13. Sato K., Azuma R., Imai T., Shimokawa T. Enhancement of mTOR signaling contributes to acquired X-ray and C-ion resistance in mouse squamous carcinoma cell line. Cancer Sci., 2017, vol. 108, no. 10, pp. 2004-2010.

14. Shimura T., Kakuda S., Ochiai Y., Nakagawa H., Kuwahara Y., Takai Y., Kobayashi J., Komatsu K., Fukumoto M. Acquired radioresistance of human tumor cells by DNA-PK/AKT/GSK3beta-mediated cyclin D1 overexpression. Oncogene, 2010, vol. 29, no. 34, pp. 4826-4837.

15. Suetens A., Moreels M., Quintens R., Soors E., Buset J., Chiriotti S., Tabury K., Gregoire V., Baatout S. Dose- and time-dependent gene expression alterations in prostate and colon cancer cells after in vitro ex-posure to carbon ion and X-irradiation. J. Radiat. Res., 2015, vol. 56, no. 1, pp. 11-21.

16. Scott J.G., Berglund A., Schell M.J., Mihaylov I., Fulp W.J., Yue B., Welsh E., Caudell J.J., Ahmed K., Strom T.S., Mellon E., Venkat P., Johnstone P., Foekens J., Lee J., Moros E., Dalton W.S., Eschrich S.A., McLeod H., Harrison L.B., Torres-Roca J.F. A genome-based model for adjusting radiotherapy dose (GARD): a retrospective, cohort-based study. Lancet Oncol., 2017, vol. 18, no. 2, pp. 202-211.

17. Khmelevsky E.V., Kancheli I.N., Khoroshkov V.S., Kaprin A.D. Morbidity dynamics in proton-photon or photon radiation therapy for locally advanced prostate cancer. Rep. Pract. Oncol. Radiother., 2018, vol. 23, no. 1, pp. 21-27.

18. Mohan R., Peeler C.R., Guan F., Bronk L., Cao W., Grosshans D.R. Radiobiological issues in proton ther-apy. Acta Oncol., 2017, vol. 56, no. 11, pp. 1367-1373.

19. Nasedkina N.V., Beketov E.E., Isaeva E.V., Malakhov E.P., Golovanova O.Y., Ulyanenko L.N., Chernu-kha A.E., Saburov V.O., Lepilina O.G., Ulyanenko S.Е. RBE of "Prometeus" facility protons for irradiation of tumor cells in vitro with one and three fields //FUTURE OF ATOMIC ENERGY – AtomFuture 2017. XIII In-ternational Youth Scientific and Practical Conference. KnE Engineering, 2018, vol. 3, pp. 309-316. Availa-ble at: https://knepublishing.com/index.php/KnE-Engineering/issue/view/80 (Accessed 03.10.2020).

20. Beketov E.E., Isaeva E.V., Nasedkina N.V., Malakhov E.P., Golovanova O.Y., Ulyanenko L.N., Chernu-kha A.E., Saburov V.O., Lepilina O.G., Ulyanenko S.E. Biological dose uniformity in spread-out Bragg peak of proton scanning beam therapeutic facility. Voprosy onkologii – Oncology Questions, 2018, vol. 64, no. 5, pp. 678-682.

21. 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.

22. Olive P.L., Banath J.P. The comet assay: a method to measure DNA damage in individual cells. Nat. Pro-toc., 2006, vol. 1, no 1, pp. 23-29.

23. Paganetti H. Relative biological effectiveness (RBE) values for proton beam therapy. Variations as a func-tion of biological endpoint, dose, and linear energy transfer. Phys. Med. Biol., 2014, vol. 59, no. 22, pp. R419-R472.

24. Culard F., Bouffard S., Charlier M. High-LET irradiation of a DNA-binding protein: protein-protein and DNA-protein crosslinks. Radiat. Res., 2005, vol. 164, no. 6, pp. 774-780.

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