Luukkonen Jukka, Höytö Anne, Sokka Miiko, Liimatainen Anu, Syväoja Juhani, Juutilainen Jukka, Naarala Jonne
a University of Eastern Finland , Department of Environmental and Biological Sciences , Yliopistonranta , Kuopio , Finland.
b University of Eastern Finland , Department of Environmental and Biological Sciences , Joensuu , Finland.
Int J Radiat Biol. 2017 Feb;93(2):240-248. doi: 10.1080/09553002.2017.1235298. Epub 2016 Oct 5.
PURPOSE: In our previous studies, exposure to extremely low frequency (ELF) magnetic fields (MF) altered responses to DNA damage caused by menadione. The aim of this study was to evaluate possible ELF MF induced changes in proteins involved in DNA damage responses and in cell cycle distribution. MATERIALS AND METHODS: Based on our previous studies, the exposure protocol included pre-exposure of human SH-SY5Y neuroblastoma cells to a 50 Hz, 100 μT MF for 24 h prior to a 3-h menadione treatment. As DNA damage responses are relatively fast processes, a 1-h menadione treatment was also included in the experiments. The menadione concentrations used were 1, 10, 15, 20, and 25 μM. Immunoblotting was used to assess the levels of DNA damage response-related proteins (γ-H2AX, Chk1, phospho-Chk1, p21, p27, and p53), while the level of DNA damage was assessed by the alkaline Comet assay. Cell cycle distribution was assayed by SYTOX Green staining followed by flow cytometry analysis. RESULTS: The main findings in MF-exposed cells were decreased p21 protein level after the 1-h menadione treatment, as well as increased proportion of cells in the G1 phase and decreased proportion of S phase cells after the 3-h menadione treatment. These effects were detectable also in the absence of menadione. CONCLUSIONS: The results indicate that MF exposure can alter the G1 checkpoint response and that the p21 protein may be involved in early responses to MF exposure.
目的:在我们之前的研究中,暴露于极低频(ELF)磁场(MF)会改变对甲萘醌引起的DNA损伤的反应。本研究的目的是评估ELF MF可能诱导的参与DNA损伤反应的蛋白质变化以及细胞周期分布的变化。 材料与方法:基于我们之前的研究,暴露方案包括在3小时甲萘醌处理之前,将人SH-SY5Y神经母细胞瘤细胞预先暴露于50 Hz、100 μT的MF中24小时。由于DNA损伤反应是相对快速的过程,实验中还包括1小时的甲萘醌处理。使用的甲萘醌浓度为1、10、15、20和25 μM。免疫印迹法用于评估DNA损伤反应相关蛋白(γ-H2AX、Chk1、磷酸化-Chk1、p21、p27和p53)的水平,而DNA损伤水平通过碱性彗星试验进行评估。细胞周期分布通过SYTOX Green染色后进行流式细胞术分析来测定。 结果:暴露于MF的细胞中的主要发现是,在1小时甲萘醌处理后p21蛋白水平降低,以及在3小时甲萘醌处理后G1期细胞比例增加,S期细胞比例降低。在没有甲萘醌的情况下也可检测到这些效应。 结论:结果表明,MF暴露可改变G1期检查点反应,并且p21蛋白可能参与对MF暴露的早期反应。
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