Analysis of radiation-induced apoptosis using radioresistant mutants

使用抗辐射突变体分析辐射诱导的细胞凋亡

基本信息

项目摘要

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.
细胞凋亡是一个重要的生理过程,可被各种物理和化学刺激如电离辐射和各种化学物质激活。为了分析辐射诱导细胞凋亡的分子机制,我们从辐射敏感的小鼠胸腺淋巴瘤3SB细胞系中分离出抗辐射突变株。在通过重复X-照射富集后,EMS处理的细胞在含有0.25%琼脂的半固体培养基中生长,并分离出12个独立的集落。将这些克隆细胞系在液体培养基中培养几周,然后在用5戈伊X射线照射后接种到半固体培养基中,以分离更稳定的抗辐射突变体。我们最终获得了5个稳定的细胞系,并发现一个克隆,1B 1C 4,比其他克隆更能抵抗X射线诱导的细胞凋亡。当3SB细胞暴露于5戈伊的X-射线时,用赤藓红B染色的细胞分数在照射后孵育8小时内迅速增加。而1B 1C 4细胞即使孵育24 h也未发生凋亡。使用琼脂糖凝胶电泳从X射线照射的1B 1C 4细胞的DNA观察到类似的辐射抗性。对p53基因第5 ~ 9外显子的cDNA片段进行PCR-SSCP分析,结果表明,1B 1C 4和1D 5 -8两个细胞克隆含有突变型p53基因。PCR直接测序结果显示,1B 1C 4细胞在p53密码子238的第二位有一个由C到T的转变。序列特异性DNA结合试验表明,这种突变导致p53蛋白的功能缺陷。这些结果表明,辐射诱导小鼠胸腺淋巴瘤细胞凋亡的过程可能至少有两条途径:p53依赖性途径和p53非依赖性途径。

项目成果

期刊论文数量(5)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
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).
K.Kobitsu 等人:“仓鼠胰管腺癌和细胞系中端粒长度缩短,端粒酶活性增加。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
T.Shimizu: "N-ras mutation detected in spontaneous neoplastic transformation of Chinese hamaster embryo cells." Tissue Culture Research Communications. 15(2). 131-140 (1996)
T.Shimizu:“在中国仓鼠胚胎细胞的自发肿瘤转化中检测到 N-ras 突变。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
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)
Y.Ishigaki:“通过将米色和 xid 突变引入 KSN 裸鼠品系培育出新的免疫缺陷小鼠品系。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
T. Shimizu: "A specific 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. 546-554 (1995)
T. Shimizu:“特定的染色体变化和独特的转化基因对于培养的中国仓鼠胚胎细胞自发转化的恶性进展是必要的”《日本癌症研究杂志》。
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
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)
T.Shimizu:“特定的染色体变化和独特的转化基因对于培养的中国仓鼠胚胎细胞自发转化的恶性进展是必要的。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

SUZUKI Fumio其他文献

An experimental approach for analysis of biological effect of low dose radiation and factors affecting DSB repair fidelity
低剂量辐射生物学效应及DSB修复保真度影响因素分析的实验方法
  • DOI:
  • 发表时间:
    2015
  • 期刊:
  • 影响因子:
    0
  • 作者:
    CAO Lili;KAWAI Hidehiko;SASATANI Megumi;IIZUKA Daisuke;MASUDA Yuji;INABA Toshiya;SUZUKI Keiji;OOTSUYAMA Akira;UMATA Toshiyuki;KAMIYA Kenji;SUZUKI Fumio;Hiroshi Tauchi
  • 通讯作者:
    Hiroshi Tauchi
The boundary between 'bad' and 'good' outsiders and the construction of unifying elements underpinning rural communities.
“坏”和“好”外来者之间的界限以及支撑农村社区的统一元素的建设。
  • DOI:
  • 发表时间:
    2012
  • 期刊:
  • 影响因子:
    0
  • 作者:
    CAO Lili;KAWAI Hidehiko;SASATANI Megumi;IIZUKA Daisuke;MASUDA Yuji;INABA Toshiya;SUZUKI Keiji;OOTSUYAMA Akira;UMATA Toshiyuki;KAMIYA Kenji;SUZUKI Fumio;加賀爪優;Shiro Horiuchi
  • 通讯作者:
    Shiro Horiuchi
耳間時間差が音像の分離知覚に与える影響
耳间时间差对声像分离知觉的影响
  • DOI:
  • 发表时间:
    2014
  • 期刊:
  • 影响因子:
    0
  • 作者:
    MASUDA Yuji;SUZUKI Miki;KAWAI Hidehiko;HISHIKI Asami;HASHIMOTO Hiroshi;MASUTANI Chikahide;HISHIDA Takashi;SUZUKI Fumio;KAMIYA Kenji;近藤成一;森川大輔
  • 通讯作者:
    森川大輔

SUZUKI Fumio的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('SUZUKI Fumio', 18)}}的其他基金

The search and analysis for essential signaling mediators responding to radiation by proteome techniques.
通过蛋白质组技术搜索和分析响应辐射的重要信号传导介质。
  • 批准号:
    17310035
  • 财政年份:
    2005
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Mechanisms of apoptotic cell death caused by radiation-induced perturbation in checkpoint regulations.
检查点调节中辐射引起的扰动引起细胞凋亡的机制。
  • 批准号:
    13480168
  • 财政年份:
    2001
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
INVESTTGATON OF FACTORS PROMOTING HIPPOCAMPAL SCLEROSIS IN THE MOUSE MODEL OF PROGRESSIVE HYPERTROPHY OF DENTATE GYRUS IN HIPPOCAMPUS.
海马齿状回进行性肥大小鼠模型中促进海马硬化的因素研究。
  • 批准号:
    13671432
  • 财政年份:
    2001
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
RESEARCH OF NOVEL GENES PROMOTING NEURONAL PLASTISITY IN ANIMAL MODEL OF HYPERTROPHIC HIPPOCAMPAL GRANULE CELLS
肥大海马颗粒细胞动物模型促进神经元可塑性的新基因研究
  • 批准号:
    10671295
  • 财政年份:
    1998
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Analysis of the checkpoint genes controlling induction of chromosome aberrations by radiation.
分析控制辐射诱导染色体畸变的检查点基因。
  • 批准号:
    10480135
  • 财政年份:
    1998
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
Induction of numerical chromosome changes and neoplastic transformation by radiations.
通过辐射诱导染色体数量变化和肿瘤转化。
  • 批准号:
    62580164
  • 财政年份:
    1987
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)

相似海外基金

What is the tissue microenvironment that alters radiation susceptibility for carcinogenesis? -Using a thymic lymphoma development model-
改变辐射致癌敏感性的组织微环境是什么?
  • 批准号:
    18K18198
  • 财政年份:
    2018
  • 资助金额:
    $ 1.47万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Role of DNA Double Strand Break Response in Suppression of Thymic Lymphoma
DNA 双链断裂反应在抑制胸腺淋巴瘤中的作用
  • 批准号:
    7780950
  • 财政年份:
    2010
  • 资助金额:
    $ 1.47万
  • 项目类别:
Molecular Pathways to Thymic Lymphoma and Leukemia
胸腺淋巴瘤和白血病的分子途径
  • 批准号:
    6828142
  • 财政年份:
    2004
  • 资助金额:
    $ 1.47万
  • 项目类别:
Molecular Pathways to Thymic Lymphoma and Leukemia
胸腺淋巴瘤和白血病的分子途径
  • 批准号:
    6935295
  • 财政年份:
    2004
  • 资助金额:
    $ 1.47万
  • 项目类别:
Molecular Pathways to Thymic Lymphoma and Leukemia
胸腺淋巴瘤和白血病的分子途径
  • 批准号:
    7122964
  • 财政年份:
    2004
  • 资助金额:
    $ 1.47万
  • 项目类别:
Molecular Pathways to Thymic Lymphoma and Leukemia
胸腺淋巴瘤和白血病的分子途径
  • 批准号:
    7250866
  • 财政年份:
    2004
  • 资助金额:
    $ 1.47万
  • 项目类别:
Molecular Pathways in T Cell Development and Thymic Lymphoma
T 细胞发育和胸腺淋巴瘤的分子途径
  • 批准号:
    6989689
  • 财政年份:
    2004
  • 资助金额:
    $ 1.47万
  • 项目类别:
Molecular Pathways to Thymic Lymphoma and Leukemia
胸腺淋巴瘤和白血病的分子途径
  • 批准号:
    7483248
  • 财政年份:
    2004
  • 资助金额:
    $ 1.47万
  • 项目类别:
P53 AND ATM CHECKPOINTS IN THYMIC LYMPHOMA SUPPRESSION
胸腺淋巴瘤抑制中的 P53 和 ATM 检查点
  • 批准号:
    6624715
  • 财政年份:
    1995
  • 资助金额:
    $ 1.47万
  • 项目类别:
P53 AND ATM CHECKPOINTS IN THYMIC LYMPHOMA SUPPRESSION
胸腺淋巴瘤抑制中的 P53 和 ATM 检查点
  • 批准号:
    6835112
  • 财政年份:
    1995
  • 资助金额:
    $ 1.47万
  • 项目类别:
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了