N-(2-butanoyloxyethyl)-4-(chloromethyl)-3-nitrobenzamide (NBCN) is a nitroaromatic bioreducible compound with cytotoxic effects in cancer cell lines. The aim of this work was to investigate the molecular mechanisms involved in cell death promoted by NBCN in HL60 cells. We observed that NBCN treatment increased intracellular ROS and reduced mitochondria membrane potential (ΔΨm). NBCN treatment also induced morphological changes, phosphatidylserine exposure, cell cycle arrest in G2/M-phase, DNA condensation and fragmentation, but it did not show cytotoxic effects on normal human peripheral blood mononuclear cells (PBMCs). NBCN-induced caspase 3- and 9-dependent DNA fragmentation, which was blocked by pretreatment with the broad-spectrum caspase inhibitor, z-VAD-fmk. Flow cytometry analysis demonstrated that NBCN also increased of the number of autophagic vesicles in HL60 cells, which was not observed when cells were pre-treated with bafilomycin A1. Taken together, these results indicate that NBCN triggered the mitochondrial apoptotic pathway and led to the onset of autophagic cell death, which contributed to its cytotoxic effects.

N-（2-丁酰氧基乙基）-4-（氯甲基）-3-硝基苯甲酰胺（NBCN）是一种硝基芳香族可生物还原的化合物，在癌细胞系中具有细胞毒性作用。这项工作的目的是研究NBCN在HL60细胞中促进细胞死亡的分子机制。我们观察到，NBCN处理可增加细胞内ROS并降低线粒体膜电位（ΔΨm）。NBCN处理还诱导了形态变化，磷脂酰丝氨酸暴露，G2 / M期细胞周期停滞，DNA浓缩和片段化，但未对正常人外周血单个核细胞（PBMC）表现出细胞毒性作用。NBCN诱导的caspase 3和9依赖性DNA断裂，被广谱caspase抑制剂z-VAD-fmk预处理阻断。流式细胞仪分析表明，NBCN也增加了HL60细胞中自噬囊泡的数量，而当细胞用bafilomycin A1预处理时未观察到。综上所述，这些结果表明NBCN触发了线粒体凋亡途径，并导致自噬细胞死亡的发生，这有助于其细胞毒性作用。