WORTMANNIN的辐射增敏与NF-kB相关  

The studies of radiosensitized effect related to NF-kB induced by WORTMANNIN

作　　者：李雨[1] 赵芳[1] 郑秀龙[1] 

机构地区：[1]第二军医大学海军医学系放射医学教研室,上海200433

出　　处：《辐射研究与辐射工艺学报》2009年第3期187-189,共3页Journal of Radiation Research and Radiation Processing

摘　　要：用凝胶迁移实验(Electrophoretic mobility shift assay,EMSA)研究PI-3K的特异性抑制剂WORTMANNIN(WT)影响细胞辐射反应与核因子NF-kB的关系。合成含kB位点的双链寡核苷酸碱基序列,以无菌双蒸水稀释,退火,制备放射性核苷酸探针。常规方法培养LP3细胞,实验LP3细胞样品受WT作用及5Gyγ射线照射后,提取核蛋白,与标记探针进行结合反应,进行非变性聚丙烯酰氨凝胶电泳。真空干胶,低温环境中进行放射自显影,照相扫描,进行转录因子的凝胶迁移率改变分析。未受WT作用的LP3细胞转录因子NF-kB结合活性在受5Gyγ射线照后6h内,经历从8%升高至50%然后降低至12%,50μmol/L WT明显抑制了转录因子受照后的变化幅度。电离辐射可能通过一些信号传导途径激活NF-kB,进而促使一些与DNA损伤修复和细胞其它防御机制有关的基因活动,以减轻辐射损伤,WT有可能是通过阻滞这一过程的早期阶段而发生辐射增敏效应。In order to investigate radiosensitized effect induced by WORTMANNIN（WT） the nuclear factor kB （NF-kB） was studied by Electrophoretic Mobility Shift Assay （EMSA）. Double strand nucleotide sequence which contains kB genetic locus was composed, diluted and annealed in pure water, and then labeled as radioactive nucleotide probe. LP3 cell was cultivated in conventional method and receive 5 Gy of γ-irradiation after nurturing with WT. The nucleoprotein picked up from the cell was combined with the radioactive labeled probe and then was carded out with electrophoresis by SDS-PAGE, which was taken photographs using autoradiograph in low temperature after vacuum drying. It has been found that the NF-kB activity of LP3 cell arises from about 8 % to 50 % and then becomes down to 12 % in 6 h after irradiation. WT at 50 μmol/L can inhibit the NF-kB activity evidently. Radiation can activate NF-kB through message pathway and promote the gene express, which is related to repair and other defense mechanism of damaged DNA in cells. WT may block the massage pathway at early stage and induce radiation sensitized effect.

关 键 词：WORTMANNIN 辐射 NF-KB 凝胶迁移实验 

分 类 号：R817.4[医药卫生—影像医学与核医学] R818[医药卫生—放射医学]