权益分类	功能权益	普通用户	{{item.name}}会员
{{category.name}}	{{benefitItem.name}}

Studies on the combination therapy of radiation and p53 gene transfer

放射与p53基因转移联合治疗的研究

基本信息

批准号：
15591316
负责人：
NAKANO Hisako
金额：
$ 1.98万
依托单位：
TOKYO MRTROPOLITAN ORGANIZATION FOR MEDICAL RESEARCH
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2004
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15591316/
关键词：
apoptosis cell death X-rays p53 MOLT-4 cells L5178Y cells M10 cells

项目摘要

DNA damage can cause cell cycle arrest or apoptosis, and both responses contribute to tumor suppression by p53. In response to DNA damage, cells with wild-type p53 genes exhibit a rapid increase in p53 protein levels. We studied the relation between individual cellular p53 level and apoptosis, and also tried to detect the induction of apoptosis on X-irradiated cells with mutant p53 in purpose to cancer therapy.Human leukemic MOLT-4 cells undergo apoptosis after X-irradiation through p53-dependent pathway. In MOLT-4 cells, p53 (wild type) is stabilized and increased after X-irradiation. We have studied to measure p53 accumulation by flow cytometry. Irradiated cells were fixed and permeabilized, stained, and analyzed on FACSClibur (Becton Dickinson). MOLT-4 cells exposed to 0.05-9.1 Gy were anlayzed for p53 at 1-6 h after the irradiation by flow cytometory. The p53 protein level was increasing by 4-5 h after the irradiation, and then decrease gradually. In the case of cells exposed to D10 or lower than, cellular p53 protein level showed that in unirradiated control cells after 24 h of postirradiation. In the other hand, cells exposed to high dose rapidly underwent apoptosis, and the cellular p53 reached the low level than control cells after 24h of postirradiation. These results show that there is the significant correlation among radiosensitivity (by colony formation), apoptosis and the amount of individual cellular p53.We also evaluated the effect of wild p53 transfer to mutant p53 expressing cells. When survival was determined by the dye-exclusion test at 24 h after irradiation, the percentage of X-ray-induced dead cells was increased.

DNA损伤可引起细胞周期阻滞或细胞凋亡，这两种反应都有助于p53抑制肿瘤。在DNA损伤的反应中，带有野生型p53基因的细胞表现出p53蛋白水平的快速增加。我们研究了单个细胞p53水平与细胞凋亡的关系，并试图检测突变p53对x射线照射细胞凋亡的诱导作用，目的是为了治疗癌症。人白血病MOLT-4细胞在x射线照射后通过p53依赖途径发生凋亡。在MOLT-4细胞中，p53（野生型）在x射线照射后稳定并升高。我们研究了用流式细胞术检测p53的积累。将辐照后的细胞固定、渗透、染色并在FACSClibur （Becton Dickinson）上进行分析。在0.05 ~ 9.1 Gy照射后1 ~ 6 h，流式细胞术检测MOLT-4细胞p53水平。照射后4 ~ 5 h， p53蛋白水平升高，随后逐渐降低。在暴露于D10或低于D10的情况下，细胞p53蛋白水平显示，在未辐照的对照细胞中，辐射后24小时。另一方面，高剂量暴露的细胞迅速发生凋亡，照射后24h细胞p53水平低于对照细胞。这些结果表明放射敏感性（通过集落形成）、细胞凋亡和单个细胞p53的数量之间存在显著相关性。我们还评估了野生p53转移到表达p53的突变细胞的效果。照射后24 h通过染料排除试验测定存活，x射线诱导的死细胞百分比增加。