The study of modulating effects on X-ray induced cell mutability by chaperons and autophagy

伴侣和自噬对X射线诱导的细胞突变的调节作用研究

基本信息

批准号：
18510041
负责人：
SUZUKI Nobuo
金额：
$ 2.64万
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：
2006
资助国家：
日本
起止时间：
2006 至 2007
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-18510041/
关键词：
annexin II aldolase A human serum chaperon autophagy X-ray human cell UV (ultraviolet)annexinII 紫外線 X線シャペロンプロテアーゼ

项目摘要

The present work is designed to investigate the role of chaperons and autophagy on the suppressive physiological functions for radiation-induced mutagenicity in human. (1) We previously reported that HSP27 is involved in cellular DNA-repair functions after UV irradiation. In the present study, annexinII was identified as a candidate gene for the HSP27 interacting protein, and suggested to be involved in cellular UV resistance by the colony forming assay using siRNA for annexinII mRNA. (2) We also tried proteomic studies for elucidation of radiation responsible genes. Using X-ray irradiated-nuclear cell extracts, we found some candidate genes. Aldolase A, one of genes for glycolytic enzyme proteins, was suggested to be involved in cellular X-ray resistance. (3) To moreover examine the involvement of autophagy in radiation response, expression levels of LC3, one of autophagy marker proteins, were analyzed using HSP27-suppressive cell lines. We could not find any difference of LC3 expression levels between HSP27-suppressive and control cells, although increased levels of LC3 was observed after serum starvation. (4) In human body oxidative stress conditions are expected to play roles on modulation of radiation response. Therefore, the test to monitor the oxidative stress by evaluating serum hydroperoxide was carried out. Amounts of serum hydroperoxide showed higher levels in patients, such as rheumatoid arthritis and diabetes mellitus, than healthy volunteers. Thus, it will be an intriguing problem whether radiation susceptibility in human is regulated by combination of chaperones metabolism with oxidative conditions.

目前的工作旨在研究伴侣和自噬对人类辐射诱导的诱导性诱变的抑制生理功能的作用。（1）我们先前报道说HSP27参与了紫外线照射后的细胞DNA修复功能。在本研究中，将末期鉴定为Hsp27相互作用蛋白的候选基因，并建议通过使用siRNA进行囊acnxinii mRNA的菌落形成分析来参与细胞UV抗性。（2）我们还尝试了蛋白质组学研究来阐明辐射负责基因。使用X射线照射的核细胞提取物，我们发现了一些候选基因。醛溶酶A是糖酵解酶蛋白的基因之一，建议参与细胞X射线抗性。（3）为了检查自噬参与辐射反应的参与，使用HSP27抑制性细胞系分析了LC3的表达水平，一种自噬标记蛋白。尽管在血清饥饿后观察到LC3水平升高，但我们找不到Hsp27抑制细胞和对照细胞之间LC3表达水平的任何差异。（4）在人体中，氧化应激条件有望在辐射反应的调节中起作用。因此，通过评估血清过氧化物来监测氧化应激的测试。与健康的志愿者相比，血清氢过氧化物的量表显示出患者（例如类风湿关节炎和糖尿病）的水平更高。因此，如果通过伴侣代谢与氧化条件的组合来调节人类中的辐射敏感性，那将是一个有趣的问题。