Yu Yun, Jia Peimin, Huang Ying, Cai Xun, Chen Guoqiang
Shanghai Institute of Hematology, Rui-Jin Hospital, Shanghai Second Medical University, Shanghai 200025, China.
Zhonghua Xue Ye Xue Za Zhi. 2002 May;23(5):254-7.
To investigate the effects of mitochondrial membrane permeability transition pore (MPT)-opened agent diamide and MPT-closed agent cyclosporin A on arsenic trioxide (As(2)O(3))-induced apoptosis in acute promyelocytic leukemia (APL) cell line NB4.
NB4 cells were treated with As(2)O(3) alone or in combination with diamide or cyclosporin A in different concentrations. Cell apoptosis was assessed by the morphological observation, Annexin-V assay, distribution of cellular DNA contents and genomic DNA electrophoresis. The mitochondrial transmembrane potentials (DeltaPsim) were detected by flow cytometry according to the intensity of rhodamine 123 uptake in cells.
Both diamide and cyclosporin A significantly enhanced As(2)O(3)-induced apoptosis in NB4 cells. The DeltaPsim collapse induced by As(2)O(3) was also enforced by combined treatment with diamide or cyclospo-rin A. 1 micromol/L As(2)O(3) alone treatment for 72 hours led to DeltaPsim disruption in 27.9% of cells, while combined treatment of As(2)O(3) and diamide or cyclosporin A increased DeltaPsim disruption cells to 59.7% and 42.2%, respectively.
As(2)O(3)-induced DeltaPsim disruption possibly involves with thiol oxidation or crosslink of important components especially ANT-related molecules.
研究线粒体膜通透性转换孔（MPT）开放剂二酰胺和MPT封闭剂环孢素A对三氧化二砷（As₂O₃）诱导急性早幼粒细胞白血病（APL）细胞系NB4凋亡的影响。
NB4细胞分别用不同浓度的As₂O₃单独处理或与二酰胺或环孢素A联合处理。通过形态学观察、膜联蛋白V检测、细胞DNA含量分布及基因组DNA电泳评估细胞凋亡。根据细胞中罗丹明123摄取强度，采用流式细胞术检测线粒体跨膜电位（ΔΨm）。
二酰胺和环孢素A均显著增强As₂O₃诱导的NB4细胞凋亡。As₂O₃诱导的ΔΨm崩溃也因与二酰胺或环孢素A联合处理而增强。单独用1 μmol/L As₂O₃处理72小时导致27.9%的细胞ΔΨm破坏，而As₂O₃与二酰胺或环孢素A联合处理使ΔΨm破坏细胞分别增加到59.7%和42.2%。
As₂O₃诱导的ΔΨm破坏可能与重要成分尤其是ANT相关分子的硫醇氧化或交联有关。
