利用拉曼光谱技术研究低剂量X射线辐射对人神经母细胞瘤细胞的影响  

Study on the Radiobiological Effects of Low-Dose X-Ray on Human Neuroblastoma Cells by Raman Spectroscopy

作　　者：陈珊 林兰 陈钧 刘樱 CHEN Shan;LIN Lan;CHEN Jun;LIU Ying(Institute of Materials,China Academy of Engineering Physics,Mianyang 621907,China;Molecular Medicine Research Center,State Key Laboratory of Biotherapy,West China Hospital,Sichuan University,Chengdu 610041,China;Science and Technology on Surface Physics and Chemistry Laboratory,Jiangyou 621908,China)

机构地区：[1]中国工程物理研究院材料研究所,四川绵阳621907 [2]四川大学华西医院生物治疗国家重点实验室分子医学中心,四川成都610041 [3]表面物理与化学重点实验室,四川江油621908

出　　处：《光谱学与光谱分析》2023年第1期129-132,共4页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(11705170,21573223);中国工程物理研究院材料研究所特聘人才基金项目(TP02201709);重点实验室稳定支撑项目(WDZC201901)资助。

摘　　要：低剂量电离辐射引发的生物效应复杂而多样,其研究往往又受到辐射标志物和检测技术手段的限制。将拉曼光谱技术应用于低剂量辐射生物效应研究,利用10 mW,532 nm共聚焦拉曼光谱对经过100,200和500 mGy三种辐射剂量的X射线辐照之后的人神经母细胞瘤细胞进行检测,发现细胞嘌呤核苷酸(722~728和1572~1581 cm^(-1)等等)、嘧啶核苷酸(770~785 cm^(-1)等等)等DNA相关的拉曼特征峰受到电离辐射影响而发生变化,说明低剂量X射线辐照造成细胞DNA水平改变。采用流式细胞术对同样条件辐照后培养6 h的人神经母细胞瘤细胞进行细胞周期分析发现,三种剂量的X射线电离辐射均造成细胞在G2期阻滞,同样提示电离辐射引起DNA水平升高。通过划痕实验分析辐照后20 h的细胞迁移能力,结果显示,相较于未接受X射线照射的对照细胞,受到三种剂量电离辐射的人神经母细胞瘤细胞均出现迁移水平下降。研究结果表明,通过拉曼光谱分析发现低剂量X射线电离辐射引起人神经母细胞瘤细胞DNA水平变化,其结果与细胞周期分析和迁移分析的结果相一致,但检测时间大大提前,利用拉曼光谱技术可以实现低剂量辐射损伤等细胞生物学效应的早期发现与监测。The biological effects induced by low-dose ionizing radiation are complex and diverse.Radiation biomarkers and detection techniques often limit the studies.In this paper,Raman spectroscopy was applied to study the biological effects of low-dose radiation.Raman spectroscopy at 10 mW and 532 nm were used to analyze human neuroblastoma cells irradiated by X-ray at 100,200 and 500 mGy.The DNA related Raman characteristic spectroscopic peaks of purine nucleotides(722~728,1572~1581 cm^(-1),etc.)and pyrimidine nucleotides(770~785 cm^(-1),etc.)were changed by ionizing radiation,indicating that low-dose X-ray irradiation caused changes in cell DNA level.Flow cytometry was used to analyze the cell cycle of human neuroblastoma cells cultured for 6 hours after irradiation under the same conditions.All three doses of X-ray ionizing radiation caused cell stagnation in the G2 phase of the cell cycle,which also suggested that ionizing radiation caused the increase of DNA level.Analysis of cell migration at 20 h after irradiation by scratch assay showed that human neuroblastoma cells exposed to all three doses of ionizing radiation showed reduced levels of migration compared to control cells not exposed to X-ray.Raman spectroscopy showed that low dose X-ray ionizing radiation induced changes in the DNA level of human neuroblastoma cells,and that was consistent with the cell cycle analysis and migration analysis,but the detection time was much earlier.Raman spectroscopy can be used to detect and monitor the early biological effects of low-dose radiation.

关 键 词：拉曼光谱 低剂量电离辐射 辐射生物效应 

分 类 号：O657.3[理学—分析化学]