1800MHz GSM移动电话射频电磁场敏感细胞筛选及生物学效应研究

发布时间：2018-05-26 16:54

本文选题：射频电磁场 + 相对敏感细胞　；参考：《浙江大学》2012年博士论文

【摘要】：移动电话射频电磁场的遗传毒性效应是其健康危险度评估中的关键科学问题。由于所采用的研究模型、辐照参数、分析方法等不同,目前有关射频电磁场遗传毒性的研究结果不仅很不一致,而且缺乏可比性,无法得出定论。有证据显示,电磁场的生物学效应与所研究的生物学系统的遗传背景相关。我们认为,在研究电磁场遗传毒性效应时,首先应该使用能指征电磁场这种弱作用因素对DNA影响的灵敏方法,系统地比较不同来源细胞对电磁场的响应情况,即确定电磁场敏感细胞；然后,以敏感细胞为研究对象,深入探索电磁场的遗传毒性和其它生物学效应。在本学位论文中,我们首先采用指示DNA双链断裂的早期、灵敏、特异的金指标一一yH2AX焦点形成分析技术,以六种不同系统来源的代表性细胞为研究对象,检测移动电话射频电磁场对DNA双链断裂的影响,以明确射频电磁场的遗传毒性作用以及该作用是否具有细胞种类依赖性；如发现敏感细胞,则以此为研究对象,进一步分析其后续生物学效应。结果显示,比吸收率(SAR)为3W/kg的1800MHz射频电磁场间断(5minon/10min off)辐照细胞1小时,没有引起中国仓鼠肺成纤维细胞(CHL)、SD大鼠星形胶质细胞(Astrocytes)、人羊膜上皮细胞(FL)、人脐静脉内皮细胞(HUVEC)、人眼晶状体上皮细胞(HLEC)和人皮肤成纤维细胞(HSF)六种细胞的DNA双链断裂；间断辐照24小时,仅观察到CHL和HSF细胞yH2AX焦点数的显著增加。在以上发现射频电磁场辐照致HSF细胞γH2AX焦点增加的基础上,我们采用彗星实验、PI染色结合细胞流式检测、细胞计数、CCK-8检测、DCFH-DA探针孵育结合细胞流式检测和免疫荧光成像等方法分析了其后续生物学效应。结果显示,射频电磁场诱导的HSF细胞γH2AX焦点增加未能引起细胞内DNA断裂片段以及细胞周期、细胞增殖和细胞活力等的显著改变；没有同步改变γH2AX焦点复合物中的53BP1和Rad51焦点的形成；也没有引起HSF细胞内ROS水平的显著变化。根据以上研究结果,我们得出如下结论：1)在本实验条件下,移动电话射频电磁场以时间相关和细胞种类依赖的方式诱导细胞γH2AX焦点的增加；2)CHL和HSF细胞是响应移动电话射频电磁场辐照的相对敏感细胞；3)移动电话射频电磁场诱导的γH2AX焦点增加并不导致显著的生物学后果。本学位论文的主要创新点：研究思路和方法方面,1)首次采用DNA损伤检测的灵敏新方法,系统评价射频电磁场的遗传毒性效应；2)首次以筛选出的射频电磁场敏感细胞为研究对象,系统分析其后续生物学效应。在科学研究方面,1)首次报道CHL和HSF细胞为射频电磁场相对敏感细胞；2)首次报道了射频电磁场诱导的γH2AX焦点增加未能引起细胞基因组不稳定等后续生物学效应的发生。
[Abstract]:Genetic toxicity of radio frequency electromagnetic field (RF EMF) in mobile phones is a key scientific problem in health risk assessment. Due to the different models, radiation parameters and analytical methods, the current research results on genetic toxicity of radiofrequency electromagnetic field are not only inconsistent, but also lack comparability, so it is impossible to draw a conclusion. There is evidence that the biological effects of electromagnetic fields are related to the genetic background of the biological systems studied. We believe that in studying the genotoxic effects of electromagnetic fields, we should first use a sensitive method that can indicate the effects of electromagnetic fields on DNA, and systematically compare the responses of cells from different sources to electromagnetic fields. That is, to determine the electromagnetic field sensitive cells, and then to explore the genotoxicity and other biological effects of the electromagnetic field by taking the sensitive cells as the research object. In this dissertation, we first used the yH2AX focus formation analysis technique, which indicates the early, sensitive and specific gold index of DNA double strand breaks, and studied the representative cells from six different systems. To detect the effects of radio frequency electromagnetic fields on DNA double strand breaks in mobile phones in order to determine the genotoxicity of radio frequency electromagnetic fields and whether they are cell type dependent and, if sensitive cells are found, to do so. Further analysis of its subsequent biological effects. The results showed that the specific absorptivity of 3W/kg was 5 min / 10 min off of the 1800MHz radiofrequency electromagnetic field (1800MHz) irradiation for 1 hour. The DNA double strand breaks of six kinds of cells, such as Astrocytes, human amniotic membrane epithelial cells, human umbilical vein endothelial cells, human ocular lens epithelial cells and human skin fibroblasts, have not been caused by the Chinese hamster lung fibroblasts (CHL) and SD rat astrocytes. After 24 hours of intermittent irradiation, only a significant increase in the number of yH2AX focal points in CHL and HSF cells was observed. On the basis of the above finding that the 纬 H2AX focus of HSF cells was increased by radiofrequency electromagnetic field irradiation, we used comet assay and Pi staining combined with flow cytometry. The subsequent biological effects were analyzed by cell counting and CCK-8 detection with DCFH-DA probe incubation combined with cell flow detection and immunofluorescence imaging. The results showed that the increase of 纬 H2AX focus in HSF cells induced by radiofrequency electromagnetic field did not cause significant changes in DNA fragmentation, cell cycle, cell proliferation and cell viability. The formation of 53BP1 and Rad51 focal points in 纬 H2AX focal complexes was not changed synchronously, nor did it cause significant changes in ROS levels in HSF cells. Based on the above results, we draw the following conclusion: 1) under the present experimental conditions, Radio frequency electromagnetic field of mobile phone induces increase of cell 纬 H2AX focal point in a time dependent and cell type dependent manner. HSF and HSF cells are relatively sensitive cells in response to radio frequency electromagnetic field irradiation of mobile phone.) Radio frequency electromagnetic field of mobile phone The induced increase in 纬 H2AX focus did not result in significant biological consequences. The main innovation of this dissertation is: research ideas and methods: for the first time, a new sensitive method of DNA damage detection is used to systematically evaluate the genotoxicity of radiofrequency electromagnetic field (RF-EMF) sensitive cells are used as the research object for the first time. The following biological effects were systematically analyzed. For the first time, we report that CHL and HSF cells are relatively sensitive to radiofrequency electromagnetic field (RF-EMF). It is the first time to report that the increase of 纬 H2AX focus induced by RF-EMF does not lead to the subsequent biological effects such as genomic instability.
【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2012
【分类号】：R363

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