Gao Fei, Yi Jing, Yuan Jing Qi, Shi Gui Ying, Tang Xue Ming
Department of Cell Biology, Shanghai Second Medical University, Shanghai 200025, China.
Cell Res. 2004 Feb;14(1):81-5. doi: 10.1038/sj.cr.7290206.
Double staining flow cytometry was performed using 7-amino actinomycin D and 6-carboxy-2', 7'-dichlorodihydrofluorescein diacetate, to detect the level fluctuation of reactive oxygen species (ROS) during the cell cycle of normal NB4 cells. Our results showed that NB4 cells possessed higher level of ROS in G2/M phase than in G1 and S phases. Double staining flow cytometry, with TdT mediated dUTP nick end labeling (Tunel) and propidium iodide (PI), indicated that As2O3 (2 microM) could induce apoptosis in NB4 cells prevailingly from G2/M phase, and this efficacy was enhanced upon co-administration of 2, 3-dimethoxy-1, 4-naphthoquinone (DMNQ) (2.5 microM) which could produce the endogenous ROS. These results suggested that different ROS level in different cell cycle phases of NB4 cells might determine the selective induction of G2/M apoptosis and the cells' susceptibility to apoptosis by As2O3.
使用7-氨基放线菌素D和6-羧基-2'，7'-二氯二氢荧光素二乙酸酯进行双染色流式细胞术，以检测正常NB4细胞在细胞周期中活性氧（ROS）水平的波动。我们的结果表明，NB4细胞在G2/M期的ROS水平高于G1期和S期。采用TdT介导的dUTP缺口末端标记法（Tunel）和碘化丙啶（PI）进行双染色流式细胞术表明，三氧化二砷（2μM）可主要从G2/M期诱导NB4细胞凋亡，并且在共同给予可产生内源性ROS的2,3-二甲氧基-1,4-萘醌（DMNQ）（2.5μM）时，这种效应会增强。这些结果表明，NB4细胞在不同细胞周期阶段的不同ROS水平可能决定了对G2/M期凋亡的选择性诱导以及细胞对三氧化二砷诱导凋亡的敏感性。
