The purpose of this study was to evaluate the combination effect of resveratrol and methoxyamine on radiosensitivity of iododeoxyuridine in spheroid culture of U87MG glioblastoma cell line using colony formation and alkaline comet assays. Spheroids on day-20 with 350 µm diameters were treated with 20 µM resveratrol and/or 6 mM methoxyamine and/or 1 µM iododeoxyuridine for one volume doubling time (67 h), and then irradiated with 2 Gy gamma-radiation (60Co) in different groups. After treatment, viability of the cells, colony forming ability and DNA damages were obtained by blue dye exclusion, colony formation and alkaline comet assay, respectively. Our results showed that methoxyamine and resveratrol could significantly reduce colony number and induce the DNA damages of glioblastoma spheroid cells treated with iododeoxyuridine in combination with gamma-rays. Therefore, methoxyamine as base excision repair inhibitor and resveratrol as hypoxia inducible factor 1-alpha inhibitor in combination with iododeoxyuridine as radiosensitizer enhanced the radiosensitization of glioblastoma spheroid cells.
ChuSHFengDFMaYBZhangHZhuZALiZQJiangPC. Promoter methylation and downregulation of SLC22A18 are associated with the development and progression of human glioma. J Trans Med2011; 9: 156–156.
2.
WeiKCHuangCYChenPYFengLYWuTWChenSMTsaiHCLuYJTsangNMTsengCKPaiPCShinJW. Evaluation of the prognostic value of CD44 in glioblastoma multiforme. Anticancer Res2010; 30: 253–9.
3.
NambiarDRajamaniPSinghRP. Effects of phytochemicals on ionization radiation-mediated carcinogenesis and cancer therapy. Mutation Res2011; 728: 139–57.
4.
HegartyTJThorntonAFDiazRFChandlerWFEnsmingerWDJunckLPageMAGebarskiSSHoodTWStetsonPL. Intra-arterial bromodeoxyuridine radiosensitization of malignant gliomas. Int J Radiat Oncol Biol Phys1990; 19: 421–8.
5.
SchulzCAMehtaMPBadieBMcGinnCJRobinsHIHayesLChappellRVolkmanJBingerKArzoomanianRSimonKAlbertiDFeierabendCTutschKDKunugiKAWildingGKinsellaTJ. Continuous 28-day iododeoxyuridine infusion and hyperfractionated accelerated radiotherapy for malignant glioma: a phase I clinical study. Int J Radiat Oncol Biol Phys2004; 59: 1107–15.
6.
WilliamsJRZhangYZhouHGridleyDSKochCJSlaterJMLittleJB. Overview of radiosensitivity of human tumor cells to low-dose-rate irradiation. Int J Radiat Oncol Biol Phys2008; 72: 909–17.
7.
Neshasteh-RizA. The effect of proliferation heterogeneity on iododeoxyuridine uptake by glioma cells cultured as monolayer and spheroids measured by flow cytometry. Cell J2000; 2: 125–79.
8.
SharmaGM. Hypoxia inducible factor-1α (HIF-1 α) and its role in tumour progression to malignancy. Online J Health Allied Sci2008; 7.
9.
BaurJASinclairDA. Therapeutic potential of resveratrol: the in vivo evidence. Nat Rev Drug Discov2006; 5: 493–506.
10.
ZhangQTangXLuQYZhangZFBrownJLeAD. Resveratrol inhibits hypoxia-induced accumulation of hypoxia-inducible factor-1alpha and VEGF expression in human tongue squamous cell carcinoma and hepatoma cells. Mol Cancer Ther2005; 4: 1465–74.
11.
TavernaSHwangHSSchuppJERadivoyevitchTSessionNNReddyGZarlingDAKinsellaTJ. Inhibition of base excision repair potentiates iododeoxyuridine-induced cytotoxicity and radiosensitization. Cancer Res2003; 63: 838–46.
12.
Neshasteh-RizABabalouiSKhoeiS. Evaluation of combination effects of 2-methoxyestradiol and methoxyamine on IUdR-induced radiosensitization in glioma spheroids. Iranian J Radiat Res2010; 7: 211–6.
13.
FrosinaG. DNA repair and resistance of gliomas to chemotherapy and radiotherapy. Mol Cancer Res2009; 7: 989–99.
14.
Neshasteh-RizASakiMKhoeiS. Cytogenetic damages from Iododeoxyuridine-induced radiosensitivity with and without Methoxyamine in human glioblastoma spheroids. Cell J2008; 10: 57–64.
15.
Kunz-SchughartLAKreutzMKnuechelR. Multicellular spheroids: a three-dimensional in vitro culture system to study tumour biology. Int J Exp Pathol1998; 79: 1–23.
16.
KhoeiSDelfanSNeshasteh-RizAMahdaviSR. Evaluation of the combined effect of 2ME2 and 60Co on the inducement of DNA damage by IUdR in a spheroid model of the U87MG glioblastoma cancer cell line using alkaline comet assay. Cell J2011; 13: 83–83.
17.
FirouziFKhoeiSMirzaeiHR. Role of resveratrol on the cytotoxic effects and DNA damages of iododeoxyuridine and megavoltage radiation in spheroid culture of U87MG glioblastoma cell line. Gen Physiol Biophys2015; 34: 43–50.
18.
FazeliGRKhoeiSNikoofarAGoliaeiB. Reduced DNA damage in tumor spheroids compared to monolayer cultures exposed to ionizing radiation. Iranian J Radiat Res2007; 5: 63–9.
19.
GalanisEBucknerJ. Chemotherapy for high-grade gliomas. Br J Cancer2000; 82: 1371–80.
20.
LacroixMAbi-SaidDFourneyDRGokaslanZLShiWDeMonteFLangFFMcCutcheonIEHassenbuschSJHollandEHessKMichaelCMillerDSawayaR. A multivariate analysis of 416 patients with glioblastoma multiforme: prognosis, extent of resection, and survival. J Neurosurg2001; 95: 190–8.
21.
EyvazzadehNNeshasteh-RizAMahdaviSRMohsenifarA. Genotoxic damage to glioblastoma cells treated with 6 MV X-radiation in the presence or absence of methoxy estradiol, IUDR or topotecan. Cell J2015; 17: 312–21.
22.
RodriguezRKinsellaTJ. Halogenated pyrimidines as radiosensitizers for high grade glioma: revisited. Int J Radiat Oncol Biol Phys1991; 21: 859–62. discussion 65.
23.
KinsellaTJMitchellJBRussoAMorstynGGlatsteinE. The use of halogenated thymidine analogs as clinical radiosensitizers: rationale, current status, and future prospects: non-hypoxic cell sensitizers. Int J Radiat Oncol Biol Phys1984; 10: 1399–406.
24.
WallaceSS. Enzymatic processing of radiation-induced free radical damage in DNA. Radiat Res1998; 150(5 Suppl): S60–79.
25.
WangYIliakisG. Effects of 5'-iododeoxyuridine on the repair of radiation induced potentially lethal damage interphase chromatin breaks and DNA double strand breaks in Chinese hamster ovary cells. Int J Radiat Oncol Biol Phys1992; 23(2): 353–60.
26.
HarrisonLBChadhaMHillRJHuKShashaD. Impact of tumor hypoxia and anemia on radiation therapy outcomes. Oncologist2002; 7: 492–508.
27.
Neshasteh-RizAREZAM. Comparison between intracerebral biodistribution of [125 I] IUdR in a rat glioma model and [125 I] IUdR uptake in spheroid multicellular culture: effect of proliferative heterogeneity. Cell J2002; 3: 159–68.
28.
KeithBSimonMC. Hypoxia-inducible factors, stem cells, and cancer. Cell2007; 129: 465–72.
FreyerJPTustanoffEFrankoAJSutherlandRM. In situ oxygen consumption rates of cells in V-79 multicellular spheroids during growth. J Cell Physiol1984; 118: 53–61.
31.
DelmasDLanconAColinDJanninBLatruffeN. Resveratrol as a chemopreventive agent: a promising molecule for fighting cancer. Curr Drug Targets2006; 7: 423–42.
32.
CalCGarbanHJazirehiAYehCMizutaniYBonavidaB. Resveratrol and cancer: chemoprevention, apoptosis, and chemo-immunosensitizing activities. Curr Med Chem Anti-cancer Agents2003; 3: 77–93.
33.
PanayiotidisMIRancourtRCPappaAWhiteCW. Effect of cell cycle growth arrest on global DNA methylation status in human lung epithelial-like (A549) cells. In vivo2006; 20: 861–5.
34.
PapadopoulouMVBloomerWD. DNA repair mechanisms are involved in the hypoxia-dependent toxicity of NLCQ-1 (NSC 709257) and its synergistic interaction with alkylating agents. In vivo2007; 21: 175–80.
35.
TurnerDPCortellinoSSchuppJECarettiELohTKinsellaTJBellacosaA. The DNA N-glycosylase MED1 exhibits preference for halogenated pyrimidines and is involved in the cytotoxicity of 5-iododeoxyuridine. Cancer Res2006; 66: 7686–93.
36.
LiuLNakatsuruYGersonSL. Base excision repair as a therapeutic target in colon cancer. Clin Cancer Res2002; 8: 2985–91.
37.
LiuzziMTalpaert-BorleM. A new approach to the study of the base-excision repair pathway using methoxyamine. J Biol Chem1985; 260: 5252–8.
38.
TavernaPLiuLHwangHSHansonAJKinsellaTJGersonSL. Methoxyamine potentiates DNA single strand breaks and double strand breaks induced by temozolomide in colon cancer cells. Mutat Res2001; 485: 269–81.
