Membrane dielectric changes indicate induced apoptosis in HL-60 cells more sensitively than surface phosphatidylserine expression or DNA fragmentation
Membrane dielectric changes indicate induced apoptosis in HL-60 cells more sensitively than surface phosphatidylserine expression or DNA fragmentation
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DOI：
10.1016/s0005-2736(02)00495-9
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发表时间：
2002-08-31
影响因子：
3.4
通讯作者：
Gascoyne, PRC
中科院分区：
文献类型：
--
作者：
Wang, XJ;Becker, FF;Gascoyne, PRC
哺乳动物细胞的比膜电容和电导率分别反映了细胞表面形态的复杂性和膜屏障的功能，已被证明对细胞的生理和病理变化作出反应。在这里，这些膜特性的培养人早幼粒细胞HL-60细胞诱导凋亡的影响进行了报道。细胞介电电泳交叉频率的测量与染料木素（GEN）处理后，推导出膜电容和电导率的变化。HL-60细胞的表观比细胞膜电容从初始值17.6 ± 0.9下降到处理后4 h的9.1 ± 0.5 mF/m2。变化开始于治疗后几分钟内，并且在磷脂酰丝氨酸（PS）的外化（如通过膜联蛋白V测定测定）和亚G1细胞亚群的出现（如通过溴化乙锭染色DNA测定）之前。用广谱半胱天冬酶抑制剂N-苄氧基羰基-Val-Ala-Asp（O-甲基）-氟甲基酮（zVAD-fetamine）处理不能阻止这些早期细胞膜介电反应，表明半胱天冬酶系统不参与。虽然膜电导率没有改变GEN治疗的第一个4小时，它上升显着，并逐步此后。最后，当屏障功能失效，细胞坏死时，它增加了许多数量级。有效的膜电容和电导率的研究结果有助于将注意力集中在膜作为早期参与细胞凋亡的位点上。结合我们以前的报告中使用的介电方法的细胞操作和分离，这些结果表明，介电泳技术应适用于快速检测，分离和定量的正常，凋亡和坏死细胞的细胞混合物。(C)2002 Elsevier Science B. V.保留所有权利。
The specific membrane capacitance and conductivity of mammalian cells, which reflect their surface morphological complexities and membrane barrier functions, respectively, have been shown to respond to cell physiologic and pathologic changes. Here, the effects of induced apoptosis on these membrane properties of cultured human promyelocytic HL-60 cells are reported. Changes in membrane capacitance and conductivity were deduced from measurements of cellular dielectrophoretic crossover frequencies following treatment with genistein (GEN). The apparent specific cell membrane capacitance of HL-60 cells fell from an initial value of 17.6 +/- 0.9 to 9.1 +/- 0.5 mF/m(2) 4 h after treatment. Changes began within minutes of treatment and preceded both the externalization of phosphatidylserine (PS), as gauged by the Annexin V assay, and the appearance of a sub-G1 cell subpopulation, as determined through ethidium bromide staining of DNA. Treatment by the broad spectrum caspase inhibitor N-benzyloxycarbony-Val-Ala-Asp(O-methyl)-fluoromethyketone (zVAD-fmk) did not prevent these early cell membrane dielectric responses, suggesting that the caspase system was not involved. Although membrane conductivity did not alter during the first 4 h of GEN treatment, it rose significantly and progressively thereafter. Finally, as the barrier function failed and the cells became necrotic, it increased by many orders of magnitude. The effective membrane capacitance and conductivity findings serve to focus attention on the membrane as a site for early participation in apoptosis. In conjunction with our prior reports of the use of dielectric methods for cell manipulation and separation, these results demonstrate that dielectrophoretic technologies should be applicable to the rapid detection, separation, and quantification of normal, apoptotic, and necrotic cells from cell mixtures. (C) 2002 Elsevier Science B.V. All rights reserved.