人绒毛膜癌细胞JAR和JEG-3感受电磁场差异性的机制研究

The potential health hazard of exposure to environmental electromagnetic fields (EMF), mainly extremely low frequency magnetic fields (ELF-MF) and radiofrequency EMF (RF-EMF), continues to cause public concern. However, the possibility of biological and health effects of exposure to EMF remains controversial and their biophysical mechanisms are unknown. Thus, it is crucial to elucidate the mechanism of cellular response to EMF exposure. In our previous study, we performed γH2AX foci formation assay, a sensitive method for detecting DNA damage, to systemically screen the cells (cell lines) sensitive to EMF exposure, and identified the human choriocarcinoma JAR and JEG-3 cells as EMF sensitive cells. Interestingly, the JAR and JEG-3 cells differentially respond to both 50 Hz magnetic fields (MF) and GSM 1800 MHz EMF exposure. To further explore the mechanism of the differential cellular response to EMF in JAR and JEG-3 cells, this study will 1) determine the threshold of EMF exposure induced injuries on cells and the threshold of cellular response to EMF with cell biology and molecular biology techniques, 2) examine the effects of 50 Hz MF and GSM 1800 MHz RF-EMF exposure on proteome and metabolome profilings in JAR and JEG-3 cells with proteomics and metabolomics approaches, and compare the diferences in proteome and metabolome between these two cells under EMF exposure, 3) identify the key proteins and metabolites involved in cellular response to EMF exposure, and analyze the relationship between the proteins and metabolites in both JAR and JEG-3 cells, 4) validate if the identified proteins and metabolites are functional in the differential cellular response to EMF exposure in both cellular system and cell free system. We expect this study to uncover the roles of key proteins and metabolites in differential cellular response to EMF exposure. Our study will shed on the biophysical mechanisms of EMF acting on organism, and provide both experimental evidence and theory supports for future health risk assessment of environmental EMF exposure.

低强度环境电磁场暴露的可能健康危害备受争议，其关键是电磁场作用引发生物学效应的作用机理不清楚。因此，了解细胞感受电磁场的机制是当前科学界的迫切任务。我们的前期研究工作表明人绒毛膜癌细胞（JAR和JEG-3）响应电磁场存在差异性，为深入研究其生物学机制，本项目拟利用细胞和分子生物学实验技术量化电磁场作用JAR和JEG-3细胞的损伤阈值和作用阈值；利用高通量蛋白质组学和代谢组学研究技术比较分析电磁场对两种细胞的蛋白质表达谱和代谢图谱的影响，筛选确定细胞响应电磁场的关键蛋白和代谢物，并整合分析蛋白和代谢物之间的关系；进一步结合细胞和非细胞系统分析关键蛋白和代谢物在细胞感受电磁场中的作用，提出电磁场作用细胞的生物学机制。项目研究预期从细胞响应电磁场的关键蛋白和代谢物角度解析细胞差异性感受电磁场的分子基础，研究成果有助于探索阐明电磁场作用的生物物理机制，为电磁场健康风险评估提供实验证据和理论依据。

低强度环境电磁场暴露的可能健康危害备受争议，其关键问题是电磁场作用引发生物学效应的作用机理不清楚。为深入探索，本项目结合细胞生物学实验、多组学技术及非细胞研究系统全面分析JAR和JEG-3细胞感受50Hz磁场和1800MHz射频电磁场差异性及机制，取得以下研究结果：1）发现两类电磁场影响细胞活力、细胞周期、DNA损伤等具有细胞依赖性和电磁场暴露频率及暴露时间依赖性，呈线性和非线性反应；2）以不同生物学终点为观察指标，电磁场暴露影响细胞行为学的作用阈值不同，分别确定了两类电磁场暴露两种细胞的作用阈值。3）发现JAR细胞和JEG-3细胞本身蛋白质表达图谱差异性决定了细胞差异性响应两类电磁场，筛选确定两种细胞共同响应50Hz磁场的关键蛋白（HSP70等），且两类电磁场均能抑制低剂量镉暴露诱导JAR细胞金属硫蛋白（MT1B、MT1F、MT2A等）的表达。4）初探了代谢组学技术应用于电磁场生物学效应及机制研究的适用性，筛选到细胞响应电磁场共性代谢途径、主要代谢物，并尝试关联电磁场暴露引起两类细胞应激响应相关信号通路及代谢途径等的共性变化。5）搭建非细胞研究系统和电磁场暴露系统整合，发现电磁场暴露引起培养液、EGF蛋白等介电常数的可逆性改变，但该效应未影响EGF与EGFR相互作用及EGF诱导的EGFR聚簇、细胞增殖等，提示电磁场可能直接作用于细胞内关键蛋白；6）结合细胞和非细胞系统分析结果，提出电磁场作用通过响应蛋白、细胞内信号通路、代谢通路、代谢物进而引起细胞行为学改变的生物学机制。项目研究从细胞响应电磁场的关键蛋白和代谢物角度初步探索细胞差异性感受电磁场的分子基础，发表基于本项目研究实验结果的论文4篇、相关论文多篇，获浙江省自然科学奖二等奖1项。研究成果为探索阐明电磁场作用的生物物理机制提供基础，为电磁场健康风险评估提供实验证据和理论依据。