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Live cell observation of EGFP expression of in vitro X-irradiated plasmid DNA in non-irradiated human cells
https://repo.qst.go.jp/records/77418
https://repo.qst.go.jp/records/77418dac52c69-5124-4644-bcd1-6713f7837760
Item type | 会議発表用資料 / Presentation(1) | |||||
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公開日 | 2019-11-13 | |||||
タイトル | ||||||
タイトル | Live cell observation of EGFP expression of in vitro X-irradiated plasmid DNA in non-irradiated human cells | |||||
タイトル | ||||||
タイトル | 非照射ヒト細胞におけるin vitro X 線照射プラスミドDNA の EGFP 発現のライブセル観察 | |||||
言語 | en | |||||
言語 | ||||||
言語 | eng | |||||
資源タイプ | ||||||
資源タイプ識別子 | http://purl.org/coar/resource_type/c_c94f | |||||
資源タイプ | conference object | |||||
アクセス権 | ||||||
アクセス権 | metadata only access | |||||
アクセス権URI | http://purl.org/coar/access_right/c_14cb | |||||
著者 |
Obata, Yui
× Obata, Yui× Hirasaki, Keishiro× Yokoya, Akinari× Akimitsu, Nobuyoshi× Obata, Yui× Hirasaki, Keishiro× Yokoya, Akinari |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | Whole cell irradiation inevitably causes various damage in not only genomic DNA but also intracellular organelles. In order to separately examine repairability of DNA damage from damage responses of organelles, EGFP-expressing plasmid DNA (in TE buffer) was exposed to X-rays in vitro, and then transfected into " NON-irradiated " human breast cancer cells (MCF7). The dynamics of EGFP fluorescence of cells was observed by a live cell imaging technique to analyze DNA repair efficiency in the cells. The EGFP expression kinetics as DNA repair efficiency was analyzed for the cells transfected with the plasmids exposed to several X-ray doses. The similar experiments were also performed using plasmids treated with nicking or restriction enzyme to obtain positive control data of SSB or DSB induction, respectively. Based on those positive control values, theoretically expected efficiencies were also calculated assuming that strand breakage of plasmid DNA by irradiation followed Poisson distribution. The numbers of cells expressing EGFP in the experimental results were considerably fewer than the theoretical values, indicating that the irradiated plasmids were less repaired in the cells. These results strongly suggest that X-irradiation causes complex types of strand break termini rather than those created by enzymatic treatments, presumably so called non-DSB type clustered DNA damage. These particular types of radiation induced damage would compromise DNA repair efficiency, consequently induce down-regulation of EGFP expression. |
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会議概要(会議名, 開催地, 会期, 主催者等) | ||||||
内容記述タイプ | Other | |||||
内容記述 | 日本放射線影響学会第62回大会 | |||||
発表年月日 | ||||||
日付 | 2019-11-14 | |||||
日付タイプ | Issued |