二硫化铝（DSF）和铜（Cu）形成的复合物对于三阴性乳腺癌（TNBC）的预防和治疗安全且有效。虽然以前的研究表明DSF/Cu诱导了铁死亡，但其机制尚不清楚。通过透射电镜观察了DSF/Cu治疗的TNBC的线粒体形态，并评估细胞内铁、脂质活性氧（ROS）、丙二醛和谷胱甘肽的水平以检测是否存在铁死亡。通过转录组测序分析检查了DSF/Cu活化的铁死亡信号通路的靶基因。通过qRT-PCR、免疫荧光和西方印迹检测了靶基因HOMX1的表达。经DSF/Cu处理24小时后，TNBC细胞的线粒体明显萎缩。添加DSF/Cu补充剂导致细胞内铁、脂质ROS和丙二醛水平显著上调，谷胱甘肽水平显著下调，这些都是铁死亡的重要标志。转录组分析证实DSF/Cu活化了铁死亡信号通路，并上调了几个与氧化还原调节特别是血红素氧合酶-1（HMOX-1）相关的铁死亡靶基因。通过添加ROS清除剂N-乙酰半胱氨酸（NAC）来抑制铁死亡，显著增加DSF/Cu处理的TNBC细胞的存活率。这些结果表明，DSF/Cu增加了脂质过氧化物并导致HMOX1活性急剧增加，从而通过铁死亡引发TNBC细胞死亡。 DSF/Cu是一种有前景的TNBC治疗药物，并可能导致基于铁死亡的人类癌症治疗策略。© 2023 The Author(s). Published by IMR Press.

The complex formed by disulfiram (DSF) and copper (Cu) is safe and effective for the prevention and treatment of triple-negative breast cancer (TNBC). Although previous studies have shown that DSF/Cu induces ferroptosis, the mechanism remains unclear.The mitochondrial morphology of TNBC treated with DSF/Cu was observed by transmission microscopy, and intracellular levels of iron, lipid reactive oxygen species (ROS), malondialdehyde, and glutathione were evaluated to detect the presence of ferroptosis. Target genes for the DSF/Cu-activated ferroptosis signaling pathway were examined by transcriptome sequencing analysis. Expression of the target gene, HOMX1, was detected by qRT-PCR, immunofluorescence and western blot.The mitochondria of TNBC cells were significantly atrophied following treatment with DSF/Cu for 24 h. Addition of DSF/Cu supplement resulted in significant up-regulation of intracellular iron, lipid ROS and malondialdehyde levels, and significant down-regulation of glutathione levels, all of which are important markers of ferroptosis. Transcriptome analysis confirmed that DSF/Cu activated the ferroptosis signaling pathway and up-regulated several ferroptosis target genes associated with redox regulation, especially heme oxygenase-1 (HMOX-1). Inhibition of ferroptosis by addition of the ROS scavenger N-acetyl-L-cysteine (NAC) significantly increased the viability of DSF/Cu-treated TNBC cells.These results show that DSF/Cu increases lipid peroxidation and causes a sharp increase in HMOX1 activity, thereby inducing TNBC cell death through ferroptosis. DSF/Cu is a promising therapeutic drug for TNBC and could lead to ferroptosis-mediated therapeutic strategies for human cancer.© 2023 The Author(s). Published by IMR Press.