The relationship between molecular mechanisms of serum factors and bystander effects modulating cell mutability.

血清因子的分子机制与旁观者效应调节细胞突变性之间的关系。

• 批准号: 14580561
• 负责人: SUZUKI Nobuo
• 金额: $ 2.3万
• 依托单位: Chiba University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2002
• 资助国家: 日本
• 起止时间: 2002 至 2003
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-14580561/
• 关键词: GENE MUTATION; HUMAN CELL; HUMAN SERUM; BYSTANDER EFFECT; CHAPERON; ULTRAVIOLET RAY; X-RAY; DNA REPAIR

We previously reported that modulation of cell mutability by human serum factors is associated with increased levels of protease activity. However, radiation-induced bystander effects are suggested to have the potential to stimulate cellular cytokines and/or reactive oxygen species. The present work is designed to examine the relationships between these two phenomena, cell mutability modulation by serum factors and bystander effects. 1.A search for modulating factors in human blood. We found that the modulating activity of serum factors on cell mutability was changed in human volunteer by Head-down-tilt bed rest (BR). Some protease activities were also changed in post-BR serum. 2.A search for cellular signaling molecules. (1) GRP94; One, of the chaperons, GRP94, was identified as a candidate gene for the mediator molecule by analysis of X-ray-induced protease. GRP94 was suggested to be involved in cellular X-ray resistance by the colony forming assay using siRNA for GRP94 mRNA. (2) We also identified the other chaperon, HSP27, by analysis of UV-induced protease. The activity of removing 6-4 photo products after UV irradiation was decreased when expression of HSP27 was. suppressed. This finding indicates that HSP27 is involved in the UV susceptibility of cells. Thus, the modulation of cell mutability by these serum factors, which may have similar molecular mechanisms to bystander effects, seems to be mediated though protease signaling by chaperons such as GRP94 and HSP27.

我们之前报道过人血清因子对细胞突变性的调节与蛋白酶活性水平的增加有关。然而，辐射引起的旁观者效应被认为有可能刺激细胞因子和/或活性氧。目前的工作旨在研究这两种现象之间的关系，即血清因子对细胞突变性的调节和旁观者效应。 1.寻找人体血液中的调节因子。我们发现，人类志愿者通过低头倾斜卧床休息（BR）改变了血清因子对细胞突变性的调节活性。 BR 后血清中的一些蛋白酶活性也发生了变化。 2.寻找细胞信号分子。 (1)GRP94；通过分析 X 射线诱导的蛋白酶，将其中一个伴侣 GRP94 确定为介体分子的候选基因。通过使用 siRNA 检测 GRP94 mRNA 的集落形成测定，表明 GRP94 参与细胞 X 射线抵抗。 (2) 我们还通过分析紫外线诱导的蛋白酶鉴定了另一个伴侣，HSP27。 HSP27表达量减少时，UV照射后去除6-4光产物的活性降低。压制。这一发现表明 HSP27 与细胞的紫外线敏感性有关。因此，这些血清因子对细胞突变性的调节可能与旁观者效应具有相似的分子机制，似乎是通过 GRP94 和 HSP27 等伴侣蛋白的蛋白酶信号传导来介导的。

项目成果

Takahashi S., et al.: "Increased levels of UV-induced protease activity in human UVAP-1 cells exposed to gravity-changing stress : involvement of E-64-sensitive proteases on suppression of UV mutagenicity"Cell Biol.Int.. 27. 53-56 (2003)

Takahashi S. 等人：“暴露于重力变化压力的人类 UVAP-1 细胞中紫外线诱导的蛋白酶活性水平增加：E-64 敏感蛋白酶参与抑制紫外线致突变性”Cell Biol.Int..

Kita K., et al.: "Involvement of Leu-13 in interferon-induced refractoriness of human RSa cells to X-ray cell killing."Radiat.Res.. 160. 302-308 (2003)

Kita K. 等人：“Leu-13 参与干扰素诱导的人 RSa 细胞对 X 射线细胞杀伤的难治性。”Radiat.Res.. 160. 302-308 (2003)

Takahashi S., et al.: "Increased levels of UV-induced protease activity in human UVAP-1 cells exposed to gravity-changing stress : involvement of E-64-sensitive proteases on suppression of UV mutagenicity"Cell Biol. Int.. (in press).

Takahashi S. 等人：“暴露于重力变化压力的人类 UVAP-1 细胞中紫外线诱导的蛋白酶活性水平增加：E-64 敏感蛋白酶参与抑制紫外线致突变性”Cell Biol。

Moriya, T., et al.: "Enhanced expression of the LDH-A gene after gravity-changing stress in human RSa cells"Biol. Sci. Space. 16. 12-17 (2002)

Moriya, T. 等人：“人 RSa 细胞中重力变化应激后 LDH-A 基因的表达增强”Biol。

Chigira S., Sugita K., Sugaya S., Arase Y., Ichinose M., Shirasawa H.Suzuki N.: "Increased expression of the huntington interacting protein-1 gene, in cells from Hutchinson Gilford syndrome (Progeria) patients and aged donors."J.Gerontology : Biological S

Chigira S.、Sugita K.、Sugaya S.、Arase Y.、Ichinose M.、Shirasawa H.Suzuki N.：“Hutchinson Gilford 综合征（早衰症）患者和 Shirasawa H.Suzuki N. 细胞中亨廷顿相互作用蛋白 1 基因的表达增加