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Open Access 01-12-2011 | Research

A nucleoside anticancer drug, 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine (TAS106), sensitizes cells to radiation by suppressing BRCA2 expression

Authors: Shunsuke Meike, Tohru Yamamori, Hironobu Yasui, Masato Eitaki, Akira Matsuda, Masami Morimatsu, Masakazu Fukushima, Yasundo Yamasaki, Osamu Inanami

Published in: Molecular Cancer | Issue 1/2011

Abstract

Background

A novel anticancer drug 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine (ECyd, TAS106) has been shown to radiosensitize tumor cells and to improve the therapeutic efficiency of X-irradiation. However, the effect of TAS106 on cellular DNA repair capacity has not been elucidated. Our aim in this study was to examine whether TAS106 modified the repair capacity of DNA double-strand breaks (DSBs) in tumor cells.

Methods

Various cultured cell lines treated with TAS106 were irradiated and then survival fraction was examined by the clonogenic survival assays. Repair of sublethal damage (SLD), which indicates DSBs repair capacity, was measured as an increase of surviving cells after split dose irradiation with an interval of incubation. To assess the effect of TAS106 on the DSBs repair activity, the time courses of γ-H2AX and 53BP1 foci formation were examined by using immunocytochemistry. The expression of DNA-repair-related proteins was also examined by Western blot analysis and semi-quantitative RT-PCR analysis.

Results

In clonogenic survival assays, pretreatment of TAS106 showed radiosensitizing effects in various cell lines. TAS106 inhibited SLD repair and delayed the disappearance of γ-H2AX and 53BP1 foci, suggesting that DSB repair occurred in A549 cells. Western blot analysis demonstrated that TAS106 down-regulated the expression of BRCA2 and Rad51, which are known as keys among DNA repair proteins in the homologous recombination (HR) pathway. Although a significant radiosensitizing effect of TAS106 was observed in the parental V79 cells, pretreatment with TAS106 did not induce any radiosensitizing effects in BRCA2-deficient V-C8 cells.

Conclusions

Our results indicate that TAS106 induces the down-regulation of BRCA2 and the subsequent abrogation of the HR pathway, leading to a radiosensitizing effect. Therefore, this study suggests that inhibition of the HR pathway may be useful to improve the therapeutic efficiency of radiotherapy for solid tumors.

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Kaina B: DNA damage-triggered apoptosis: critical role of DNA repair, double-strand breaks, cell proliferation and signaling. Biochem Pharmacol. 2003, 66: 1547-1554. 10.1016/S0006-2952(03)00510-0CrossRefPubMed

Castedo M, Perfettini JL, Roumier T, Andreau K, Medema R, Kroemer G: Cell death by mitotic catastrophe: a molecular definition. Oncogene. 2004, 23: 2825-2837. 10.1038/sj.onc.1207528CrossRefPubMed

Mirzayans R, Severin D, Murray D: Relationship between DNA double-strand break rejoining and cell survival after exposure to ionizing radiation in human fibroblast strains with differing ATM/p53 status: implications for evaluation of clinical radiosensitivity. Int J Radiat Oncol Biol Phys. 2006, 66: 1498-1505. 10.1016/j.ijrobp.2006.08.064CrossRefPubMed

Shrivastav M, De Haro LP, Nickoloff JA: Regulation of DNA double-strand break repair pathway choice. Cell Res. 2008, 18: 134-147. 10.1038/cr.2007.111CrossRefPubMed

Stark JM, Hu P, Pierce AJ, Moynahan ME, Ellis N, Jasin M: ATP hydrolysis by mammalian RAD51 has a key role during homology-directed DNA repair. J Biol Chem. 2002, 277: 20185-20194. 10.1074/jbc.M112132200CrossRefPubMed

Baumann P, West SC: Role of the human RAD51 protein in homologous recombination and double-stranded-break repair. Trends Biochem Sci. 1998, 23: 247-251. 10.1016/S0968-0004(98)01232-8CrossRefPubMed

Shivji MK, Davies OR, Savill JM, Bates DL, Pellegrini L, Venkitaraman AR: A region of human BRCA2 containing multiple BRC repeats promotes RAD51-mediated strand exchange. Nucleic Acids Res. 2006, 34: 4000-4011. 10.1093/nar/gkl505PubMedCentralCrossRefPubMed

Esashi F, Christ N, Gannon J, Liu Y, Hunt T, Jasin M, West SC: CDK-dependent phosphorylation of BRCA2 as a regulatory mechanism for recombinational repair. Nature. 2005, 434: 598-604. 10.1038/nature03404CrossRefPubMed

Tutt A, Gabriel A, Bertwistle D, Connor F, Paterson H, Peacock J, Ross G, Ashworth A: Absence of Brca2 causes genome instability by chromosome breakage and loss associated with centrosome amplification. Curr Biol. 1999, 9: 1107-1110. 10.1016/S0960-9822(99)80479-5CrossRefPubMed

10.

Verhaegh GW, Jongmans W, Morolli B, Jaspers NG, van der Schans GP, Lohman PH, Zdzienicka MZ: A novel type of X-ray-sensitive Chinese hamster cell mutant with radioresistant DNA synthesis and hampered DNA double-strand break repair. Mutat Res. 1995, 337: 119-129.CrossRefPubMed

11.

Hattori H, Tanaka M, Fukushima M, Sasaki T, Matsuda A: Nucleosides and nucleotides. 158. 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)-cytosine, 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)uracil, and their nucleobase analogues as new potential multifunctional antitumor nucleosides with a broad spectrum of activity. J Med Chem. 1996, 39: 5005-5011. 10.1021/jm960537gCrossRefPubMed

12.

Shimamoto Y, Koizumi K, Okabe H, Kazuno H, Murakami Y, Nakagawa F, Matsuda A, Sasaki T, Fukushima M: Sensitivity of human cancer cells to the new anticancer ribo-nucleoside TAS-106 is correlated with expression of uridine-cytidine kinase 2. Jpn J Cancer Res. 2002, 93: 825-833.CrossRefPubMed

13.

Matsuda A, Sasaki T: Antitumor activity of sugar-modified cytosine nucleosides. Cancer Sci. 2004, 95: 105-111. 10.1111/j.1349-7006.2004.tb03189.xCrossRefPubMed

14.

Naito T, Yokogawa T, Kim HS, Matsuda A, Sasaki T, Fukushima M, Wataya Y: An apoptotic pathway of 3'-Ethynylcytidine (ECyd) involving the inhibition of RNA synthesis mediated by RNase L. Nucleic Acids Res Suppl. 2006, 50: 103-104.CrossRef

15.

Koizumi K, Shimamoto Y, Azuma A, Wataya Y, Matsuda A, Sasaki T, Fukushima M: Cloning and expression of uridine/cytidine kinase cDNA from human fibrosarcoma cells. Int J Mol Med. 2001, 8: 273-278.PubMed

16.

Maehara Y, Nakamura H, Nakane Y, Kawai K, Okamoto M, Nagayama S, Shirasaka T, Fujii S: Activities of various enzymes of pyrimidine nucleotide and DNA syntheses in normal and neoplastic human tissues. Gann. 1982, 73: 289-298.PubMed

17.

Inanami O, Iizuka D, Iwahara A, Yamamori T, Kon Y, Asanuma T, Matsuda A, Kashiwakura I, Kitazato K, Kuwabara M: A novel anticancer ribonucleoside, 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine, enhances radiation-induced cell death in tumor cells. Radiat Res. 2004, 162: 635-645. 10.1667/RR3268CrossRefPubMed

18.

Yasui H, Inanami O, Asanuma T, Iizuka D, Nakajima T, Kon Y, Matsuda A, Kuwabara M: Treatment combining X-irradiation and a ribonucleoside anticancer drug, TAS106, effectively suppresses the growth of tumor cells transplanted in mice. Int J Radiat Oncol Biol Phys. 2007, 68: 218-228. 10.1016/j.ijrobp.2006.12.061CrossRefPubMed

19.

Yasui H, Ogura A, Asanuma T, Matsuda A, Kashiwakura I, Kuwabara M, Inanami O: Inhibition of HIF-1alpha by the anticancer drug TAS106 enhances X-ray-induced apoptosis in vitro and in vivo. Br J Cancer. 2008, 99: 1442-1452. 10.1038/sj.bjc.6604720PubMedCentralCrossRefPubMed

20.

Hall EJ: Repair of Radiation Damage and the Dose-Rate Effect. Radiobiology for the radiologist. Edited by: Hall EJ, Giaccia AJ. 2006, Philadelphia: Lippincott Willams & Wilkins, 6

21.

Rogakou EP, Boon C, Redon C, Bonner WM: Megabase chromatin domains involved in DNA double-strand breaks in vivo. J Cell Biol. 1999, 146: 905-916. 10.1083/jcb.146.5.905PubMedCentralCrossRefPubMed

22.

Schultz LB, Chehab NH, Malikzay A, Halazonetis TD: p53 binding protein 1 (53BP1) is an early participant in the cellular response to DNA double-strand breaks. J Cell Biol. 2000, 151: 1381-1390. 10.1083/jcb.151.7.1381PubMedCentralCrossRefPubMed

23.

Kraakman-van der Zwet M, Overkamp WJ, van Lange RE, Essers J, van Duijn-Goedhart A, Wiggers I, Swaminathan S, van Buul PP, Errami A, Tan RT, Jaspers NG, Sharan SK, Kanaar R, Zdzienicka MZ: Brca2 (XRCC11) deficiency results in radioresistant DNA synthesis and a higher frequency of spontaneous deletions. Mol Cell Biol. 2002, 22: 669-679. 10.1128/MCB.22.2.669-679.2002PubMedCentralCrossRefPubMed

24.

Hickson I, Zhao Y, Richardson CJ, Green SJ, Martin NM, Orr AI, Reaper PM, Jackson SP, Curtin NJ, Smith GC: Identification and characterization of a novel and specific inhibitor of the ataxia-telangiectasia mutated kinase ATM. Cancer Res. 2004, 64: 9152-9159. 10.1158/0008-5472.CAN-04-2727CrossRefPubMed

25.

Wachters FM, van Putten JW, Maring JG, Zdzienicka MZ, Groen HJ, Kampinga HH: Selective targeting of homologous DNA recombination repair by gemcitabine. Int J Radiat Oncol Biol Phys. 2003, 57: 553-562. 10.1016/S0360-3016(03)00503-0CrossRefPubMed

26.

Takagi M, Sakata K, Someya M, Tauchi H, Iijima K, Matsumoto Y, Torigoe T, Takahashi A, Hareyama M, Fukushima M: Gimeracil sensitizes cells to radiation via inhibition of homologous recombination. Radiother Oncol. 2010, 96: 259-266. 10.1016/j.radonc.2010.05.020CrossRefPubMed

27.

Utsumi H, Elkind MM: Requirement for repair of DNA double-strand breaks by homologous recombination in split-dose recovery. Radiat Res. 2001, 155: 680-686. 10.1667/0033-7587(2001)155[0680:RFRODD]2.0.CO;2CrossRefPubMed

28.

Rao BS, Tano K, Takeda S, Utsumi H: Split dose recovery studies using homologous recombination deficient gene knockout chicken B lymphocyte cells. J Radiat Res (Tokyo). 2007, 48: 77-85. 10.1269/jrr.06050CrossRef

29.

Collis SJ, Tighe A, Scott SD, Roberts SA, Hendry JH, Margison GP: Ribozyme minigene-mediated RAD51 down-regulation increases radiosensitivity of human prostate cancer cells. Nucleic Acids Res. 2001, 29: 1534-1538. 10.1093/nar/29.7.1534PubMedCentralCrossRefPubMed

30.

Chan N, Koritzinsky M, Zhao H, Bindra R, Glazer PM, Powell S, Belmaaza A, Wouters B, Bristow RG: Chronic hypoxia decreases synthesis of homologous recombination proteins to offset chemoresistance and radioresistance. Cancer Res. 2008, 68: 605-614. 10.1158/0008-5472.CAN-07-5472CrossRefPubMed

31.

Yu D, Sekine E, Fujimori A, Ochiya T, Okayasu R: Down regulation of BRCA2 causes radio-sensitization of human tumor cells in vitro and in vivo. Cancer Sci. 2008, 99: 810-815. 10.1111/j.1349-7006.2008.00741.xCrossRefPubMed

32.

Lee SA, Roques C, Magwood AC, Masson JY, Baker MD: Recovery of deficient homologous recombination in Brca2-depleted mouse cells by wild-type Rad51 expression. DNA Repair (Amst). 2009, 8: 170-181. 10.1016/j.dnarep.2008.10.002CrossRef

33.

Brown ET, Holt JT: Rad51 overexpression rescues radiation resistance in BRCA2-defective cancer cells. Mol Carcinog. 2009, 48: 105-109. 10.1002/mc.20463PubMedCentralCrossRefPubMed

34.

Tabata S, Tanaka M, Endo Y, Obata T, Matsuda A, Sasaki T: Anti-tumor mechanisms of 3'-ethynyluridine and 3'-ethynylcytidine as RNA synthesis inhibitors: development and characterization of 3'-ethynyluridine-resistant cells. Cancer Lett. 1997, 24 (116): 225-231.CrossRef

35.

Shimamoto Y, Kazuno H, Murakami Y, Azuma A, Koizumi K, Matsuda A, Sasaki T, Fukushima M: Cellular and biochemical mechanisms of the resistance of human cancer cells to a new anticancer ribo-nucleoside, TAS-106. Jpn J Cancer Res. 2002, 93: 445-452.CrossRefPubMed

36.

Azuma A, Matsuda A, Sasaki T, Fukushima M: 1-(3-C-ethynyl-beta-D-ribo-pentofuranosyl)cytosine (ECyd, TAS-106)1: antitumor effect and mechanism of action. Nucleosides Nucleotides Nucleic Acids. 2001, 20: 609-619. 10.1081/NCN-100002337CrossRefPubMed

Title: A nucleoside anticancer drug, 1-(3-C-ethynyl-β-D-ribo-pentofuranosyl)cytosine (TAS106), sensitizes cells to radiation by suppressing BRCA2 expression
Authors: Shunsuke Meike
Tohru Yamamori
Hironobu Yasui
Masato Eitaki
Akira Matsuda
Masami Morimatsu
Masakazu Fukushima
Yasundo Yamasaki
Osamu Inanami
Publication date: 01-12-2011
Publisher: BioMed Central
Published in: Molecular Cancer / Issue 1/2011
Electronic ISSN: 1476-4598
DOI: https://doi.org/10.1186/1476-4598-10-92

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