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In vitro studies of DNA damage and repair mechanisms induced by BNCT in a poorly differentiated thyroid carcinoma cell line

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Abstract

Boron neutron capture therapy (BNCT) for aggressive tumors is based on nuclear reaction [10B (n, α) 7Li]. Previously, we demonstrated that BNCT could be applied for the treatment of undifferentiated thyroid carcinoma. The aim of the present study was to describe the DNA damage pattern and the repair pathways that are activated by BNCT in thyroid cells. We analyzed γH2AX foci and the expression of Ku70, Rad51 and Rad54, main effector enzymes of non-homologous end joining (NHEJ) and homologous recombination repair (HRR) pathways, respectively, in thyroid follicular carcinoma cells. The studied groups were: (1) C [no irradiation], (2) gamma [60Co source], (3) N [neutron beam alone], (4) BNCT [neutron beam plus 10 µg 10B/ml of boronphenylalanine (10BPA)]. The total absorbed dose was always 3 Gy. The results showed that the number of nuclear γH2AX foci was higher in the gamma group than in the N and BNCT groups (30 min–24 h) (p < 0.001). However, the focus size was significantly larger in BNCT compared to other groups (p < 0.01). The analysis of repair enzymes showed a significant increase in Rad51 and Rad54 mRNA at 4 and 6 h, respectively; in both N and BNCT groups and the expression of Ku70 did not show significant differences between groups. These findings are consistent with an activation of HRR mechanism in thyroid cells. A melanoma cell line showed different DNA damage pattern and activation of both repair pathways. These results will allow us to evaluate different blocking points, to potentiate the damage induced by BNCT.

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References

  • Aihara T, Hiratsuka J, Morita N, Uno M, Sakurai Y, Maruhashi A, Ono K, Harada T (2006) First clinical case of boron neutron capture therapy for head and neck malignancies using 18F-BPA PET. Head Neck 28(9):850–855

    Article  Google Scholar 

  • Aiyama H, Nakai K, Yamamoto T, Nariai T, Kumada H, Ishikawa E, Isobe T, Endo K, Takada T, Yoshida F, Shibata Y, Matsumura A (2011) A clinical trial protocol for second line treatment of malignant brain tumors with BNCT at University of Tsukuba. Appl Radiat Isot 69(12):1819–1822

    Article  Google Scholar 

  • Antonelli F, Campa A, Esposito G, Giardullo P, Belli M, Dini V, Meschini S, Simone G, Sorrentino E, Gerardi S, Cirrone GA, Tabocchini MA. (2015). Induction and Repair of DNA DSB as Revealed by H2AX Phosphorylation Foci in Human Fibroblasts Exposed to Low- and High-LET radiation: relationship with early and delayed reproductive cell death. Radiat Res 183(4):417–431. https://doi.org/10.1667/RR13855.1

  • Belli M, Sapora O, Tabocchini MA (2002) Molecular targets in cellular response to ionizing radiation and implications in space radiation protection. J Radiat Res 43:13–19

    Article  Google Scholar 

  • Bracalente C, Ibañez I, Molinari B, Palmieri M, Kreiner A, Valda A, Davidson J, Durán H (2013). Induction and persistence of large γH2AX foci by high linear energy transfer radiation in DNA-dependent protein kinase-deficient cells. Int J Radiat Oncol Biol Phys 87(4):785–794

  • Burma S, Chen BP, Murphy M, Kurimasa A, Chen DJ (2001) ATM phosphorylates histone H2AX in response to DNA double-strand breaks. J Biol Chem 276(45):42462–42467

    Article  Google Scholar 

  • Busse PM, Harling OK, Palmer MR, Kiger WS 3rd, Kaplan J, Kaplan I, Chuang CF, Goorley JT, Riley KJ, Newton TH, Santa Cruz GA, Lu XQ, Zamenhof RG (2003) A critical examination of the results from the Harvard-MIT NCT program phase I clinical trial of neutron capture therapy for intracranial disease. J Neurooncol 62(1–2):111–121

  • Chiacchio S, Lorenzoni A, Boni G, Rubello D, Elisei R, Mariani G (2008) Anaplastic thyroid cancer: prevalence, diagnosis and treatment. Minerva Endocrinol 33(4):341–357

    Google Scholar 

  • Coderre JA, Morris GM (1999) The radiation biology of Boron Neutron Capture Therapy. Radiat Res 151(1):1–18

    Article  ADS  Google Scholar 

  • Dagrosa MA, Viaggi M, Kreimann E, Farías S, Garavaglia R, Agote M, Cabrini RL, Dadino JL, Juvenal GJ, Pisarev MA (2002) Selective uptake of p-borophenylalanine by undifferentiated thyroid carcinoma for boron neutron capture therapy. Thyroid 12(1):7–12

    Article  Google Scholar 

  • Dagrosa MA, Viaggi M, Longhino J, Calzetta O, Cabrini R, Edreira M, Juvenal G, Pisarev MA (2003) Experimental application of boron neutron capture therapy to undifferentiated thyroid carcinoma. Int J Radiat Oncol Biol Phys 57(4):1084–1092

    Article  Google Scholar 

  • Dagrosa MA, Thomasz L, Longhino J, Perona M, Calzetta O, Blaumann H, Rebagliati RJ, Cabrini R, Kahl S, Juvenal GJ, Pisarev MA (2007) Optimization of boron neutron capture therapy for the treatment of undifferentiated thyroid cancer. Int J Radiat Oncol Biol Phys 69(4):1059–1066

    Article  Google Scholar 

  • Dagrosa A, Carpano M, Perona M, Thomasz L, Nievas S, Cabrini R, Juvenal G, Pisarev M (2011a) Studies for the application of boron neutron capture therapy to the treatment of differentiated thyroid cancer. Appl Radiat Isot 69(12):1752–1755

    Article  Google Scholar 

  • Dagrosa MA, Crivello M, Perona M, Thorp S, Santa Cruz GA, Pozzi E, Casal M, Thomasz L, Cabrini R, Kahl S, Juvenal GJ, Pisarev MA (2011b) First evaluation of the biologic effectiveness factors of boron neutron capture therapy (BNCT) in a human colon carcinoma cell line. Int J Radiat Oncol Biol Phys 79(1):262–268

    Article  Google Scholar 

  • Dudás A, Chovanec M (2004) DNA double-strand break repair by homologous recombination. Mutat Res 566(2):131–167

    Article  Google Scholar 

  • Guerra L, Bover L, Mordoh J (1990) Differentiating effect of L-tyrosine on the human melanoma cells line IIB-MEL. Exp Cell Res 188(1):61–65

    Article  Google Scholar 

  • Ibañez I, Bracalante C, Molinari B, Palmieri M, Policastro L, Kreiner A, Burlon A, Valda A, Navalesi D, Davidson J, Davidson M, Vazquez M, Ozafrán M, Durán H (2009). Induction and rejoining of DNA double strand breaks assessed by H2AX phosphorylation in melanoma cells irradiated with proton and lithium beams. Int J Radiat Oncol Biol Phys, 74(4):1226–1235

    Article  Google Scholar 

  • Jeggo P, Löbrich M (2006) Radiation-induced DNA damage responses. Radiat Prot Dosimetry 122(1–4):124–127

    Article  Google Scholar 

  • Jen-Chung Ko Shih-CiCiou, Cheng C-M, Wang L-H, Hong J-H, Jheng M-Y, Ling S, Lin Y (2008) Involvement of Rad51 in cytotoxicity induced by epidermal growth factor receptor inhibitor (gefitinib, IressaR) and chemotherapeutic agents in human lung cancer cells. Carcinogenesis 29(7):1448–1458

  • Joensuu H, Kankaanranta L, Tenhunen M, Saarilahti K (2011) Boron Neutron Capture Therapy (BNCT) as cancer treatment. Duodecim 127(16):1697–1703

    Google Scholar 

  • Kim JS, Krasieva TB, Kurumizaka H, Chen DJ, Taylor AM, Yokomori K (2005) Independent and sequential recruitment of NHEJ and HR factors to DNA damage sites in mammalian cells. J Cell Biol 170(3):341–347

    Article  Google Scholar 

  • Korabiowska M, Quentin T, Schlott T, Bauer H, Kunze E (2004) Down-regulation of Ku 70 Ku 80 mRNA expression in transitional cell carcinomas of the urinary bladder related to tumor progression. World J Urol 2(6):431–440

  • Lieber MR (2010) The mechanism of double-strand DNA break repair by the non homologous DNA end-joining pathway. Annu Rev Biochem 79:181–211

    Article  Google Scholar 

  • Maier P, Hartmann L, Wenz F, Herskind C (2016) Cellular pathways in response to ionizing radiation and their targetability for tumor radiosensitization. Int J Mol Sci. https://doi.org/10.3390/ijms17010102

    Google Scholar 

  • Matsuda M, Miyagawa K, Takahashi M, Fukuda T, Kataoka T, Asahara T, Inui H, Watatani M, Yasutomi M, Kamada N, Dohi K, Kamiya K (1999) Mutations in the RAD54 recombination gene in primary cancers. Oncogene 18:3427–3430

  • Miller M, Quintana J, Ojeda J, Langan S, Thorp S, Pozzi E, Sztejnberg M, Estryk G, Nosal R, Saire E, Agrazar H, Graiño F (2009) New irradiation facility for biomedical applications at the RA-3 reactor thermal column. Appl Radiat Isot 67(7–8 Suppl):S226-9

    Google Scholar 

  • Olive PL (1998) The role of DNA single- and double-strand breaks in cell killing by ionizing radiation. Radiat Res 150(5 Suppl):S42–S51

    Article  ADS  Google Scholar 

  • Pasieka JL (2003) Anaplastic thyroid cancer. Curr Opin Oncol 15(1):78–83

    Article  Google Scholar 

  • Pawlik TM, Keyomarsi K (2004) Role of cell cycle in mediating sensitivity to radiotherapy. Int J Radiat Oncol Biol Phys 59(4):928–942

    Article  Google Scholar 

  • Perona M, Rodríguez C, Carpano M, Thomasz L, Nievas S, Olivera M, Thorp S, Curotto P, Pozzi E, Kahl S, Pisarev M, Juvenal G, Dagrosa A (2013). Improvement of the Boron Neutron Capture therapy (BNCT) by the previous administration of the histone deacetylase inhibitor sodium butyrate for the treatment of thyroid carcinoma. Radiat Environ Biophys. 52(3):363–373

  • Redon CE, Nakamura AJ, Zhang YW (2010) Histone gamma H2AX and poly (ADP-ribose) as clinical pharmacodynamics biomarkers. Clin Cancer Res, 16:4532–4542

    Article  Google Scholar 

  • Sigurdsson S, Van Komen S, Petukhova G, Sung P (2002) Homologous DNA pairing by human recombination factors Rad51 and Rad54. J Biol Chem 277(45):42790–42794

    Article  Google Scholar 

  • Sonoda E, Hochegger H, Saberi A, Taniguchi Y, Takeda S (2006) Differential usage of non-homologous end joining and homologous recombination in double strand break repair. DNA Rep 5:1021–1029

    Article  Google Scholar 

  • Staaf E, Brehwens K, Haghdoost S, Czub J, Wojcik A (2012) Gamma-H2AX foci in cells exposed to a mixed beam of X-rays and alpha particles. Genome Integrity 3(1):8. https://doi.org/10.1186/2041-9414-3-8

    Article  Google Scholar 

  • Vermeulen C, Verwijs-Janssen M, Begg AC, Vens C (2008) Cell cycle phase dependent role of DNA polymerase beta in DNA repair and survival after ionizing radiation. Radiother Oncol 86(3):391–398

    Article  Google Scholar 

  • Wang H, Zhang X, Wang P, Yu X, Essers J, Chen D, Kanaar R, Takeda S, Wang Y (2010) Characteristics of DNA-binding proteins determine the biological sensitivity to high-linear energy transfer radiation. Nucleic Acids Res 38(10):3245–3251

    Article  Google Scholar 

  • Weterings E, Chen DJ (2008) The endless tale of non-homologous end-joining. Cell Res 18:114–124

    Article  Google Scholar 

  • Yano K, Morotomi-Yano K, Adachi N, Akiyama H (2009) Molecular mechanism of protein assembly on DNA double-strand breaks in the non-homologous end-joining pathway. J Radiat Res 50(2):97–108

    Article  Google Scholar 

Download references

Acknowledgements

A part of these studies was supported by grants from the Scientific and Technical Research National Council (CONICET) and Secretary of Science and Technology (SEPCYT).

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Correspondence to M. A. Dagrosa.

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Rodriguez, C., Carpano, M., Curotto, P. et al. In vitro studies of DNA damage and repair mechanisms induced by BNCT in a poorly differentiated thyroid carcinoma cell line. Radiat Environ Biophys 57, 143–152 (2018). https://doi.org/10.1007/s00411-017-0729-y

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  • DOI: https://doi.org/10.1007/s00411-017-0729-y

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