突然変異細胞を用いた放射線によるアポトーシス誘 導機構の解析
著者 鈴木 文男
著者別表示 Suzuki Fumio
雑誌名 平成8(1996)年度 科学研究費補助金 基盤研究(C) 研究成果報告書概要
巻 1995 1996
ページ 3p.
発行年 1999‑03‑08
URL http://doi.org/10.24517/00066292
Creative Commons : 表示 ‑ 非営利 ‑ 改変禁止 http://creativecommons.org/licenses/by‑nc‑nd/3.0/deed.ja
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1996 Fiscal Year Final Research Report Summary
Analysis of radiation-induced apoptosis using radioresistant mutants
Research Project
Project/Area Number
07680576
Research Category
Grant-in-Aid for Scientific Research (C)
Allocation Type
Single-year Grants
Section
⼀般
Research Field
環境影響評価(含放射線⽣物学)
Research Institution
HIROSHIMA UNIVERSITY (1996) Kanazawa University (1995)
Principal Investigator
SUZUKI Fumio Res.Inst.Radiat.Biol.Med.HIROSHIMA UNIVERSITY Professor, 原爆放射能医学研究所, 教授 (10019672)
Co-Investigator(Kenkyū-buntansha)
ISHIGAKI Yasuhito Kanazawa Univ., Facul.Pharm.Sci.Associate Researcher, 薬学部, 助⼿ (20232275)
Project Period (FY)
1995 – 1996
Keywords
Mouse / Thymic lymphoma / X-rays / Mutant / Apoptosis / Radioresistant / PCR-SSCP analysis / p53 gene
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(12 results)All Other All Publications (12 results)
URL: https://kaken.nii.ac.jp/report/KAKENHI-PROJECT-07680576/076805761996kenkyu_seika_hokoku_
Research Abstract
Apoptosis is an important physiological process that can be activated by various physical and chemical stimuli such as ionizing radiation and various chemicals. In order to analyze molecular mechanism of radiation-induced apoptosis, we isolated apoptosis-resistant mutants from a radiosensitive mouse thymic lymphoma 3SB cell line following treatment with ethyl methanesulfonate (EMS). After enrichment by repeated X-irradiation, EMS treated cells were grown in a semi-solid medium containing 0.25% agar, and 12 independent colonies were isolated. These clonal cell lines were cultured in a liquid medium for a few weeks and then inoculated into the semi-solid medium following irradiation with 5 Gy X-rays, to isolate more stable radioresistant mutants. We finally obtained 5 stable cell lines and found that one clone, 1B1C4, is more resistant to X-ray-induced apoptotic cell death than other clones. When 3SB cells were exposed to 5 Gy of X-rays, the fraction of cells stained with erythrosin B increased quickly within 8 h of incubation following irradiation. However, no such apoptosis occurred in 1B1C4 cells, even after incubation for 24 h. Similar radioresistance was observed using agarose gel electrophoresis of DNA from X-irradiated 1B1C4 cells. PCR-SSCP analysis of p53 cDNA fragments containing exons 5 to 9 suggested that two clones, 1B1C4 and 1D5-8 cells, contain a mutant p53 gene. PCR direct sequence analysis revealed that 1B1C4 cells have a transition from C to T in the second position of p53 codon 238. This mutation was found to cause a functional defect in the p53 protein as revealed by the sequence-specific DNA binding assay. These results indicate that the process of radiation-induced apoptosis in the mouse thymic lymphoma cell line may take at least two routs, p53-dependent or -independent pathways.
[Publications] T.Shimizu: "A speciffic chromosome change and distinctive transforming genes are necessary for malignant progression of spontaneous transformation in cultured Chinese hamster embryo cells." Japanese Journal of Cancer Research. 86(6). 546-554 (1995)
[Publications] T.Shimizu: "N-ras mutation detected in spontaneous neoplastic transformation of Chinese hamaster embryo cells." Tissue Culture
Research Communications. 15(2). 131-140 (1996)
[Publications] Y.Ishigaki: "New immunodeficient mouse strains bred by introducing beige and xid mutations into KSN nude strain." Laboratory Animal
Science. 46(4). 418-424 (1996)
[Publications] F.Suzuki: "p53 mutation and neoplastic transformation in cultured hamster embryo cells." Japanese Journal of Veterinary Reserarch.
44(4). 244-245 (1997)
[Publications] K.Kobitsu: "Shortened telomere length and increased telomerase activity in hamster pancreatic duct adenocarcinomas and cell lines"
Molecular Carcinogenesis. 18(3)(in press). (1997)
[Publications] 鈴⽊ ⽂男: "DNA鎖切断を検出する⽅法(抗変異原・抗発がん物質とその検出法)" 講談社サイエンティフィク(⿊⽥⾏昭 編), 12 (1995)
[Publications] 鈴⽊ ⽂男: "細胞⽣物学実験法・細胞レベルでの実験(細胞解析法 I)" 広川書店(⼤熊勝治 編)(印刷中), (1997)
[Publications] T.Shimizu et al.: "A specific chromosome change and distinctive transforming genes are necessary for malignant progression of
spontaneous transformation in cultured Chinese hamster embryo cells." Jap.J.Cancer Res.86(6). 546-554 (1995)
[Publications] T.Shimizu et al.: "N-ras mutation detected in spontaneous transformation of Chinese hamster embryo cells."
Tiss.Cult.Res.Commun.15(2). 131-140 (1996)
[Publications] Y.Ishigaki et al.: "New immunodeficient mouse strains bred by introducing beige and xid mutations into KSN nude strain." Lab.Animal
Sci.46(4). 418-424 (1996)
[Publications] F.Suzuki: "p53 mutation and neoplastic transformation in cultured hamster embryo cells." Jap.J.Veter.Res.44(4). 244-245 (1997) [Publications] K.Kobitsu et al.: "Shortened telomere length and increased telomerase activity in hamster pancreatic duct adenocarcinomas and cell
lines." Mol.Carcinog.18(3), (in press).
Published: 1999-03-08
