Analysis of signal transduction and nuclear denaturation in radiation-induced cell death

辐射诱导细胞死亡中的信号转导和核变性分析

• 批准号: 12470185
• 负责人: SUZUKI Norio
• 金额: $ 1.15万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470185/
• 关键词: Ionizing radiation-induced apoptosis; SAPK; JNK; Ceramide; MOLT-4; p41 specific antibody; MOLT-4細胞; 放射線細胞死; p53; p41

The overall goal of the present project has been to analyze a mechanism of X-ray-induced apoptotic cell death in human T-cell leukemia cell line MOLT-4. Toward this goal, the purpose of the project were 1) to examine the involvement of p53 and/or SAPK/JNK, 2) to identify genes downstream of the p53 or SAPK/JNK, and to analyze the expressions of apoptosis-related genes by quantative RT-PCR methods, 3) to study a role of a new radiation-induced protein p41 in cell death.1) To examine the involvement of ceramide-SAPK/JNK pathway, the effect of acid sphingomyelinase inhibitor (D609) on X-ray-induced apoptotic cell death was studied. D609 suppressed X-ray-induced apoptotic cell death as well as phosphorylation of JNK.2) RT-PCR analysis of apoptosis-related genes revealed that the expression of c-myc gene was reduced following X-irradiation, but not in an X-ray-resistant variant cell line Rh-1a. The introduction of c-myc antisense oligonucleotides into MOLT-4 cells or the treatment of c-Myc inhibitor induced cell death.3) Specific antibodies for p41 and p42 have been generated. Western blot analysis showed that the activation of caspase-9, caspase-7, and caspas-3 ran paralleled with the generation of p41. Especially, caspase-7 among those was considered important in p41 generation.

本项目的总体目标是分析X射线诱导人T细胞白血病细胞系MOLT-4细胞凋亡的机制。为此，本项目的目的是：1)研究P53和/或SAPK/JNK的参与；2)确定P53或SAPK/JNK下游的基因，并用定量RT-PCR方法分析凋亡相关基因的表达；3)研究一种新的辐射诱导蛋白p41在细胞死亡中的作用。1)为了研究神经酰胺-SAPK/JNK通路的参与，研究酸性鞘磷脂酶抑制剂(D609)对X射线诱导的细胞死亡的影响。D609抑制X射线诱导的细胞死亡和JNK2的磷酸化。2)RT-PCR分析显示，X射线照射后c-myc基因表达降低，但对X射线抗性变异细胞系Rh-1a的影响不大。将c-myc反义寡核苷酸导入MOLT-4细胞或c-Myc抑制剂治疗诱导的细胞死亡。3)p41和p42的特异性抗体产生。Western印迹分析表明，caspase-9、caspase-7和caspas-3的激活与p41的产生是平行的。其中，caspase-7被认为在p41的生成中起重要作用。

项目成果

Ishii, T. et al.: "Brief Note and Evaluation of Acute-Radiation Syndrome and Treatment of a Tokaimura Criticality Accident Patient"J. Radit. Res.. 42. S167-S182 (2001)

Ishii, T. 等人：“急性辐射综合症的简要说明和评估以及东海村危重事故患者的治疗”J。

Enomoto, A., Suzuki, N., Liu, C., Kang, Y., Zun, J., Serizawa, S., Mastumoto, Y., Morita, A., Ito, M. and Hosi, Y.: "Involvement of c-Jun NH_2-terminal kinase-1 in heat-induced apoptotic cell death of human monoblastic leukemia U937 cells"Int. J. Radiat.

Enomoto, A.、Suzuki, N.、Liu, C.、Kang, Y.、Zun, J.、Serizawa, S.、Mastumoto, Y.、Morita, A.、Ito, M. 和 Hosi, Y.：

Kang,Y.: "Increased expression after X-irradiation of MUC1 in cultured human colon carcinoma HT-29 cells."Jpn.J.Cancer Res.. 91. 324-330 (2000)

Kang,Y.：“培养的人结肠癌 HT-29 细胞中 MUC1 的 X 射线照射后表达增加。”Jpn.J.Cancer Res.. 91. 324-330 (2000)

Hosoi, Y., et al.: "Phosphorothioate oligonucleotides, suramin and heparin inhibit DNA-dependent protein kinase activity"British Journal of Cancer. (in press).

Hosoi，Y.等人：“硫代磷酸酯寡核苷酸、苏拉明和肝素抑制DNA依赖性蛋白激酶活性”英国癌症杂志。

Mori, N. et al.: "Variations in Prkdc encoding the catalytic subunit of DNA-dependent protein kinase(DNA-PKcs)and susceptibility to radiation-induced apoptosis and lymphomagenesis"Oncogene. 20. 3609-3619 (2001)

Mori, N. 等人：“编码 DNA 依赖性蛋白激酶 (DNA-PKcs) 催化亚基的 Prkdc 的变异以及对辐射诱导的细胞凋亡和淋巴瘤发生的易感性”癌基因。