Differential effects in cells exposed to ultra-short, high intensity electric fields: cell survival, DNA damage, and cell cycle analysis
Differential effects in cells exposed to ultra-short, high intensity electric fields: cell survival, DNA damage, and cell cycle analysis
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DOI：
10.1016/j.mrgentox.2003.08.006
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发表时间：
2003-12-09
影响因子：
1.9
通讯作者：
Buescher, S
中科院分区：
文献类型：
--
作者：
Stacey, M;Stickley, J;Buescher, S
高功率、纳秒脉冲电场(NsPEF)效应一直被关注于细菌的去污，但它对哺乳动物细胞的影响现在正在被揭示。在nsPEF施加期间，使用高达300KV/cm的电场幅度向电池施加持续时间为10、60或300 ns的电脉冲。由于超短的脉冲持续时间，传递到电池的能量可以忽略不计，只观察到非热效应。我们研究了nsPEF对不同来源和生长特性的10个人细胞系和1个小鼠细胞系的遗传毒性。我们提供的数据检验了nsPEF暴露对细胞存活的影响，通过克隆形成或活细胞计数来评估；通过彗星试验和染色体畸变来确定DNA损伤；以及通过测量暴露细胞的有丝分裂指数来确定细胞周期参数。使用这些指标，我们观察到不同细胞类型之间的差异效应，非贴壁细胞对nsPEF暴露的遗传毒性效应比贴壁细胞更敏感。非贴壁培养显示细胞活力迅速下降(90%)，诱导DNA损伤，在60 ns脉冲和60kV/cm的电场作用下，达到有丝分裂的细胞数量减少。在贴壁培养的细胞中没有观察到这些影响，除了小鼠3T3细胞株，它表现出与非贴壁培养相似的生存特征。这些数据表明，nsPEF的遗传毒性可能是细胞类型特有的，因此在选择性地从一种细胞类型中去除另一种细胞类型方面具有潜在的应用，例如，在疾病状态下。(C)2003爱思唯尔B.V.保留所有权利。
High power, nanosecond pulsed electric field (nsPEF) effects have been focused on bacterial decontamination, but the impact on mammalian cells is now being revealed. During nsPEF applications, electrical pulses of 10, 60 or 300 ns durations were applied to cells using electric field amplitudes as high as 300kV/cm. Because of the ultra-short pulse durations, the energy transferred to cells is negligible, and only non-thermal effects are observed. We investigated the genotoxicity of nsPEF on adherent and non-adherent cell lines including 10 human lines and one mouse cell line with different origin and growth characteristics. We present data examining the effects of nsPEF exposure on cell survival assessed by clonogenic formation or live cell count; DNA damage determined by the comet assay and chromosome aberrations; and cell cycle parameters by measuring the mitotic indices of exposed cells. Using each of these indicators, we observed differential effects among cell types with non-adherent cells being more sensitive to the genotoxic effects of nsPEF exposures than adherent cells. Non-adherent cultures showed a rapid decrease in cell viability (90%), induction of DNA damage, and a decrease in the number of cells reaching mitosis after one 60 ns pulse with an electric field intensity of 60 kV/cm. These effects were not observed in cells grown as adherent cultures, with the exception of the mouse 3T3 cell line, which showed survival characteristics similar to non-adherent cultures. These data suggest that nsPEF genotoxicity may be cell type specific, and therefore have potential applications in the selective removal of one cell type from another, for example, in diseased states. (C) 2003 Elsevier B.V. All rights reserved.