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Journal Articles

Live-cell imaging study of mitochondrial morphology in mammalian cells exposed to X-rays

Noguchi, Miho; Kanari, Yukiko; Yokoya, Akinari; Narita, Ayumi; Fujii, Kentaro

Radiation Protection Dosimetry, 166(1-4), p.101 - 103, 2015/09

 Times Cited Count:0 Percentile:100(Environmental Sciences)

Journal Articles

Visualisation of cell cycle modifications by X-ray irradiation of single HeLa cells using fluorescent ubiquitination-based cell cycle indicators

Kaminaga, Kiichi; Noguchi, Miho; Narita, Ayumi; Sakamoto, Yuka; Kanari, Yukiko; Yokoya, Akinari

Radiation Protection Dosimetry, 166(1-4), p.91 - 94, 2015/09

 Times Cited Count:5 Percentile:44.04(Environmental Sciences)

Journal Articles

Real-time observation of irradiated HeLa-cell modified by fluorescent ubiquitination-based cell-cycle indicator using synchrotron X-ray microbeam

Narita, Ayumi; Kaminaga, Kiichi; Yokoya, Akinari; Noguchi, Miho; Kobayashi, Katsumi*; Usami, Noriko*; Fujii, Kentaro

Radiation Protection Dosimetry, 166(1-4), p.192 - 196, 2015/09

 Times Cited Count:2 Percentile:73.04(Environmental Sciences)

For the knowledge about irradiation effects of mammalian cells depending on the cell cycle, most of them had been analyzed by statistical approches. Our purpose is to establish the method for real-time observation of irradiated cells under a microscope. Fluorescent ubiquitination-based cell cycle indicator (FUCCI) human cancer (HeLa) cells (red indicates G1; green, S/G2) were exposed to a synchrotron X-ray microbeam. Cells in either G1 or S/G2 were selectively irradiated according to cell color observed in the same microscopic field in a culture dish. Time-lapse micrographs of the irradiated cells were acquired for 24 h after irradiation. The cell cycle was strongly arrested by irradiation at S/G2 and never progressed to G1. In contrast, cells irradiated at G1 progress to S/G2 with a similar time course as non-irradiated control cells. These results show single FUCCI cell exposure and live cell imaging are powerful methods for studying radiation effects on the cell cycle.

Journal Articles

Live-cell imaging study of mitochondrial morphology in mammalian cells irradiated

Kanari, Yukiko; Noguchi, Miho; Kaminaga, Kiichi; Sakamoto, Yuka; Yokoya, Akinari

Journal of Radiation Research, 55(Suppl.1), p.i129 - i130, 2014/03

Journal Articles

Visualization of cell-cycle modification by ionizing irradiation in single HeLa cells using fluorescent ubiquitination-based cell-cycle indicator

Kaminaga, Kiichi; Sakamoto, Yuka; Kanari, Yukiko; Noguchi, Miho; Yokoya, Akinari

Journal of Radiation Research, 55(Suppl.1), p.i127 - i128, 2014/03

Journal Articles

Three-dimensional culture of HeLa-FUCCI cells for study of bystander cell cycle effect of high LET particles

Sakamoto, Yuka; Kaminaga, Kiichi; Kanari, Yukiko; Noguchi, Miho; Yokoya, Akinari

Journal of Radiation Research, 55(Suppl.1), p.i120 - i121, 2014/03

Journal Articles

Significance of DNA Polymerase I in ${it in vivo}$ processing of clustered DNA damage

Shikazono, Naoya; Akamatsu, Ken; Takahashi, Momoko*; Noguchi, Miho; Urushibara, Ayumi; O'Neill, P.*; Yokoya, Akinari

Mutation Research; Fundamental and Molecular Mechanisms of Mutagenesis, 749(1-2), p.9 - 15, 2013/09

 Times Cited Count:9 Percentile:60.59(Biotechnology & Applied Microbiology)

We examined the biological consequences of bi-stranded clustered damage sites, consisting of a combination of DNA lesions using a bacterial plasmid-based assay. The transformation efficiencies were significantly lower for the bi-stranded clustered GAP/AP lesions than for either a single GAP or a single AP site. When the two lesions were separated by 10-20 bp, the transformation efficiencies were comparable with those of the single lesions. This recovery of transformation efficiency for separated lesions requires DNA polymerase I (Pol I) activity. Analogously, the mutation frequency was enhanced in a bi-stranded cluster containing a GAP and an 8-oxoG, and Pol I was found to play an important role in minimising mutations induced as a result of clustered lesions. These results indicate that the biological consequences of clustered DNA damage strongly depend on Pol I activity.

Journal Articles

Evaluation of SCCVII tumor cell survival in clamped and non-clamped solid tumors exposed to carbon-ion beams in comparison to X-rays

Hirayama, Ryoichi*; Uzawa, Akiko*; Takase, Nobuhiro*; Matsumoto, Yoshitaka*; Noguchi, Miho; Koda, Kana*; Ozaki, Masakuni*; Yamashita, Kei*; Li, H.*; Kase, Yuki*; et al.

Mutation Research; Genetic Toxicology And Environmental Mutagenesis, 756(1-2), p.146 - 151, 2013/08

 Times Cited Count:14 Percentile:45.32(Biotechnology & Applied Microbiology)

Journal Articles

The Mutagenic potential of 8-oxoG/single strand break-containing clusters depends on their relative positions

Noguchi, Miho; Urushibara, Ayumi; Yokoya, Akinari; O'Neill, P.*; Shikazono, Naoya

Mutation Research; Fundamental and Molecular Mechanisms of Mutagenesis, 732(1-2), p.34 - 42, 2012/04

 Times Cited Count:11 Percentile:56.87(Biotechnology & Applied Microbiology)

The effect of a single strand break associated with base lesion(s) in vivo remains largely unknown. In the present study we determined the mutagenicities of two- and three-lesion clustered damage sites containing a 1-nucleotide gap (GAP) and 8-oxo-7,8-dihydroguanine(s) (8-oxoG(s)). The mutation frequencies (MFs) of bi-stranded two-lesion clusters (GAP/8-oxoG), especially in mutY-deficient strains, were high and were similar to those for bi-stranded clusters with 8-oxoG and base lesions/AP sites, suggesting that the GAP is processed with an efficiency similar to the efficiency of processing a base lesion or an AP site within a cluster. The MFs of tandem two-lesion clusters comprised of a GAP and an 8-oxoG were comparable to or less than the MF of a single 8-oxoG. The mutagenic potential of three-lesion clusters, which were comprised of a tandem lesion (a GAP and an 8-oxoG) and an opposing single 8-oxoG, was higher than that of a single 8-oxoG, but was no more than that of a bi-stranded 8-oxoGs. We suggest that incorporation of a nucleotide opposite 8-oxoG is less mutagenic when a GAP is present in a cluster than when a GAP is absent. Our observations indicate that the repair of a GAP is retarded by an opposing 8-oxoG, but not by a tandem 8-oxoG, and that the extent of GAP repair determines the biological consequences.

Journal Articles

A Novel technique using DNA denatureation to detect multiplly induced single-strand breaks in a hydrated plasmid DNA molecule by X-ray and $$^{4}$$He$$^{2+}$$ ion irradiation

Yokoya, Akinari; Shikazono, Naoya; Fujii, Kentaro; Noguchi, Miho; Urushibara, Ayumi

Radiation Protection Dosimetry, 143(2-4), p.219 - 225, 2011/02

 Times Cited Count:3 Percentile:68.76(Environmental Sciences)

Multiple single-strand breaks (m-SSBs), which are predicted to be preferentially induced by high LET radiation, would be underestimated if one uses the conventional method using plasmid DNA, because m-SSBs will not cause additionally conformational changes if they are on the same or on the opposite strand but separated each other sufficiently so as not to induce a double strand break. In order to observe the invisible m-SSBs, we have developed a novel technique using DNA denaturation. The m-SSBs arising in both strands of DNA are revealed as molecular size change in single strand DNA (SS-DNA) by gel electrophoresis. We have applied this method to the X- and He$$^{2+}$$ ion irradiated sample of hydrated pUC18 plasmid DNA. A half of SS-DNA population remains as intact within the experimental resolution ($$<$$ 140 bases) for both irradiations. Contrary to our initial expectation, these results indicate that SSBs are not multiply induced over 140 bp even by high-LET irradiation.

Journal Articles

A Model for analysis of the yield and the level of clustering of radiation-induced DNA-strand breaks in hydrated plasmids

Shikazono, Naoya; Yokoya, Akinari; Urushibara, Ayumi; Noguchi, Miho; Fujii, Kentaro

Radiation Protection Dosimetry, 143(2-4), p.181 - 185, 2011/02

 Times Cited Count:2 Percentile:77.73(Environmental Sciences)

Journal Articles

Induction of DNA DSB and its rejoining in clamped and non-clamped tumours after exposure to carbon ion beams in comparison to X-rays

Hirayama, Ryoichi*; Uzawa, Akiko*; Matsumoto, Yoshitaka*; Noguchi, Miho; Kase, Yuki*; Takase, Nobuhiro*; Ito, Atsushi*; Koike, Sachiko*; Ando, Koichi*; Okayasu, Ryuichi*; et al.

Radiation Protection Dosimetry, 143(2-4), p.508 - 512, 2011/02

 Times Cited Count:9 Percentile:33.97(Environmental Sciences)

We studied double-strand breaks (DSB) induction and rejoining in clamped and non-clamped transplanted tumours in mice leg after exposure to 80 keV/$$mu$$m carbon ions and X-rays. The yields of DSB in the tumours were analysed by a static-field gel electrophoresis. The OER of DSB after X-rays was 1.68, and this value was not changed after 1 h rejoining time (1.40). These damages in oxygenated conditions were rejoined 60-70% within 1 h in situ. No difference was found between the exposure to X-rays and carbon ions for the induction and rejoining of DSB. Thus, the values of OER and rejoined fraction after exposure to carbon ions were similar to those after X-rays, and the calculated relative biological effectivenesses of carbon ion were around 1 under both oxygen conditions. The yields of DSB in vivo depend on exposure doses, oxygen conditions and rejoining time, but not on the types of radiation quality.

Journal Articles

Recent progress in the energy recovery linac project in Japan

Sakanaka, Shogo*; Akemoto, Mitsuo*; Aoto, Tomohiro*; Arakawa, Dai*; Asaoka, Seiji*; Enomoto, Atsushi*; Fukuda, Shigeki*; Furukawa, Kazuro*; Furuya, Takaaki*; Haga, Kaiichi*; et al.

Proceedings of 1st International Particle Accelerator Conference (IPAC '10) (Internet), p.2338 - 2340, 2010/05

Future synchrotron light source using a 5-GeV energy recovery linac (ERL) is under proposal by our Japanese collaboration team, and we are conducting R&D efforts for that. We are developing high-brightness DC photocathode guns, two types of cryomodules for both injector and main superconducting (SC) linacs, and 1.3 GHz high CW-power RF sources. We are also constructing the Compact ERL (cERL) for demonstrating the recirculation of low-emittance, high-current beams using above-mentioned critical technologies.

Journal Articles

Enhanced radiation-induced cell killing by Herbimycin A pre-treatment

Noguchi, Miho; Hirayama, Ryoichi*; Druzhinin, S.*; Okayasu, Ryuichi*

Radiation Physics and Chemistry, 78(12), p.1184 - 1187, 2009/12

 Times Cited Count:4 Percentile:65.27(Chemistry, Physical)

Herbimycin A (HA), as in Geldanamycin, binds to conserved pockets of heat shock protein 90 (Hsp90) and inhibits its chaperone functions. Hsp90 plays an integral role in cancer cell growth and survival, because it maintains the stability of several key proteins by its chaperone's activity. It is known that some of the proteins associated with radiation responses are functionally stabilized by Hsp90. In this study, we investigated the effect of HA on radiosensitivity in human cancer cells and the mechanism related to the sensitization. In order to gain a mechanistic insight of this sensitization, we examined repair of DNA double strand breaks (DSBs) in irradiated human cancer cells pre-treated with HA, as unrepaired DSBs are thought to be the main cause of radiation-induced cell death. Cellular radiosensitivity was determined by clonogenic assay, and the DSB rejoining kinetics was examined by constant field gel electrophoresis. SQ-5, a lung squamous carcinoma cell line, showed synergistic increase in radiosensitivity when cells were pre-treated with HA. In addition, HA significantly inhibited repair of radiation induced DSBs. These results suggest that the combination of HA and ionizing radiation may be a useful therapeutic strategy for treating certain cancer cells.

Journal Articles

Radioprotection by DMSO in nitrogen-saturated mammalian cells exposed to helium ion beams

Hirayama, Ryoichi*; Matsumoto, Yoshitaka*; Kase, Yuki*; Noguchi, Miho; Ando, Koichi*; Ito, Atsushi*; Okayasu, Ryuichi*; Furusawa, Yoshiya*

Radiation Physics and Chemistry, 78(12), p.1175 - 1178, 2009/12

 Times Cited Count:8 Percentile:44.61(Chemistry, Physical)

The contribution of OH radical-mediated indirect action by particle beams under hypoxic irradiation condition was investigated by using a radical scavenger. V79 cells were irradiated with 150 MeV/nucleon helium ions at an LET of 2.2 keV/mm in the presence or absence of DMSO, and their colony survivals were determined. The contribution of indirect action to cell killing under hypoxic condition was estimated to be 52 %. We conclude that OH radical mediated indirect action still has a half in total contribution on cell killing under hypoxic condition.

Journal Articles

Contributions of direct and indirect actions in cell killing by high-LET radiations

Hirayama, Ryoichi*; Ito, Atsushi*; Tomita, Masanori*; Tsukada, Teruyo*; Yatagai, Fumio*; Noguchi, Miho; Matsumoto, Yoshitaka*; Kase, Yuki*; Ando, Koichi*; Okayasu, Ryuichi*; et al.

Radiation Research, 171(2), p.212 - 218, 2009/02

 Times Cited Count:79 Percentile:6.63(Biology)

The biological effects of radiation originate principally in damages to DNA. DNA damages by X-rays as well as heavy ions are induced by a combination of direct and indirect actions. The contribution of indirect action in cell killing can be estimated from the maximum degree of protection by dimethylsulfoxide (DMSO), which suppresses indirect action without affecting direct action. Exponentially growing Chinese hamster V79 cells were exposed to high-LET radiations of 20 to 2106 keV/$$mu$$m in the presence or absence of DMSO and their survival was determined using a colony formation assay. The contribution of indirect action to cell killing decreased with increasing LET. However, the contribution did not reach zero even at very high LETs and was estimated to be 32% at an LET of 2106 keV/$$mu$$m. Therefore, even though the radiochemically estimated G value of OH radicals was nearly zero at an LET of 1000 keV/$$mu$$m, indirect action by OH radicals contributed to a substantial fraction of the biological effects of high-LET radiations. The RBE determined at a survival level of 10% increased with LET, reaching a maximum value of 2.88 at 200 keV/$$mu$$m, and decreased thereafter. When the RBE was estimated separately for direct action (RBE(D)) and indirect action (RBE(I)); both exhibited an LET dependence similar to that of the RBE, peaking at 200 keV/$$mu$$m. However, the peak value was much higher for RBE(D) (5.99) than RBE(I) (1.89). Thus direct action contributes more to the high RBE of high-LET radiations than indirect action does.

Journal Articles

The Yield, processing, and biological consequences of clustered DNA damage induced by ionizing radiation

Shikazono, Naoya; Noguchi, Miho; Fujii, Kentaro; Urushibara, Ayumi*; Yokoya, Akinari

Journal of Radiation Research, 50(1), p.27 - 36, 2009/01

 Times Cited Count:88 Percentile:12.91(Biology)

After living cells are exposed to ionizing radiation, a variety of chemical modifications of DNA are induced either directly by ionization of DNA or indirectly through interactions with water-derived radicals. Clustered DNA damage, which is defined as two or more of such lesions within one to two helical turns of DNA induced by a single radiation track, is considered to be a unique feature of ionizing radiation. A double strand break (DSB) is a type of clustered DNA damage. Formation and repair of DSBs have been studied in great detail over the years as they have been linked to important biological endpoints. Although non-DSB clustered DNA damage has received less attention, there is growing evidence of its biological significance. This review focuses on the current understanding of (1) the yield of non-DSB clustered damage induced by ionizing radiation (2) the processing, and (3) biological consequences of non-DSB clustered DNA damage.

Journal Articles

Progress in R&D efforts on the energy recovery linac in Japan

Sakanaka, Shogo*; Ago, Tomonori*; Enomoto, Atsushi*; Fukuda, Shigeki*; Furukawa, Kazuro*; Furuya, Takaaki*; Haga, Kaiichi*; Harada, Kentaro*; Hiramatsu, Shigenori*; Honda, Toru*; et al.

Proceedings of 11th European Particle Accelerator Conference (EPAC '08) (CD-ROM), p.205 - 207, 2008/06

Future synchrotron light sources based on the energy-recovery linacs (ERLs) are expected to be capable of producing super-brilliant and/or ultra-short pulses of synchrotron radiation. Our Japanese collaboration team is making efforts for realizing an ERL-based hard X-ray source. We report recent progress in our R&D efforts.

Oral presentation

17-allylamino-17-demethoxygeldanamycin enhances the cytotoxicity of tumor cells irradiated with carbon ions

Noguchi, Miho; Hirayama, Ryoichi*; Okayasu, Ryuichi*

no journal, , 

We investigated radiosensitization effect and its mechanism of Hsp90 inhibitor 17-AAG in human tumor cell lines irradiated with high LET carbon ions. Human tumor cell lines, DU145 derived from prostate carcinoma and normal human fibroblasts HFL III were incubated for 24 h in the presence of 17-AAG at concentration of 100nM. The cells were then irradiated with carbon ions (290MeV/nucleon, LET70keV/um) and several biological endpoints were compared. Cellular radiation sensitivity was determined by clonogenic assay and DNA double strand break (DSB) repair kinetics were examined by constant field gel electrophoresis. DU145 cells showed an increase in carbon ions-induced cell death when pre-treated with 17-AAG. The radiosensitivity enhancement ratios measured at a survival rate of 10% were 2.13 for DU145 cells. In contrast to the tumor cell lines, normal human fibroblasts with carbon irradiation showed no radiosensitization with 17-AAG pre-treatment. Our constant field gel electrophoresis studies indicated that 17-AAG had almost no effect on carbon ion-induced DSB repair in DU145 cells. On the other hand, radiation induced Rad51 foci formation showed different kinetics between the carbon ion alone and the combined treatment with 17-AAG and carbon ions in DU145 cells. Our findings suggest that mechanisms other than inhibition of DSB repair could be involved with the radiosensitization by 17-AAG in tumor cells irradiated with carbon ions. However, limited inhibition of homologous recombination by this agent may still be a possibility.

Oral presentation

The Heat shock protein 90 inhibitor 17-AAG may cause DNA double strand break repair inhibition

Noguchi, Miho; Yu, D.*; Hirayama, Ryoichi*; Kubota, Nobuo*; Okayasu, Ryuichi*

no journal, , 

The aim of this study is to evaluate the radiosensitization of Hsp90 inhibitor 17-Allylamino-17-demethoxygeldanamycin (17AAG), specifically the effect of 17AAG on the DNA DSB repair machinery. Our constant field gel electrophoresis studies indicated that pretreatment with 17AAG for 24 hours inhibited radiation induced DSB repair in two cancer cell lines (DU145 and SQ-5). The treatment of 17AAG alone leads to the reduction of Rad51 protein expression by western blotting, and the combined treatment with X-irradiation caused a delay in the formation of nuclear Rad51 foci by immuno-staining. These results suggest that 17AAG affects the key protein(s) for HRR, resulting in the radiosensitization of tumor cells. Our data show for the first time that 17AAG is a DNA DSB repair inhibitor, predominantly affecting the homologous recombination pathway.

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