The anti-tumour effect of Huaier has been demonstrated in a variety of tumours. Ferroptosis is a newly identified type of programmed cell death accompanied by the accumulation of reactive oxygen species (ROS) and iron in cells and plays a key role in the therapeutic process against malignant tumours. We aimed to explore the potential therapeutic role of Huaier in pancreatic cancer and uncover the relationship between Huaier and ferroptosis.

CCK8 and colony formation assays were used to determine the proliferation of pancreatic cancer cells (PCs). The levels of cellular ROS were analysed by a fluorescence probe, and the accumulation of cellular iron was showed by Prussian blue staining. The autophagosomes and mitochondrial morphology were characterised by transmission electron microscopy (TEM). The levels of intracellular glutathione (GSH) and lipid peroxidation were measured by the corresponding kits.

The growth inhibitory effect of Huaier on PCs was concentration- and time-dependent, but this effect was significantly attenuated by ferroptosis inhibitors. In addition, Huaier effectively inhibited the GSH–GPX4 antioxidation system and resulted in the massive accumulation of ROS in PCs As shown by TEM, Huaier-treated PCs exhibited a decrease in mitochondrial cristae and a smaller mitochondrion, accompanied by an increase in autophagosomes. Indeed, we found that autophagy can induce ferroptosis in PCs and that Huaier-induced ferroptosis can be suppressed by the autophagosome inhibitor, Wortmannin.

Huaier can activate ferroptosis by inducing autophagy in PCs.

槐耳的抗肿瘤作用已在多种肿瘤中得到证实。铁死亡是一种新发现的程序性细胞死亡类型，伴随着细胞内活性氧（ROS）和铁的积累，在恶性肿瘤的治疗过程中起着关键作用。我们旨在探索槐耳在胰腺癌中的潜在治疗作用，并揭示槐耳与铁死亡之间的关系。

CCK8 和集落形成测定用于确定胰腺癌细胞 (PC) 的增殖。通过荧光探针分析细胞活性氧的水平，并通过普鲁士蓝染色显示细胞铁的积累。通过透射电子显微镜（TEM）表征自噬体和线粒体形态。通过相应的试剂盒测量细胞内谷胱甘肽（GSH）和脂质过氧化的水平。

槐耳对 PCs 的生长抑制作用呈浓度和时间依赖性，但这种作用被铁死亡抑制剂显着减弱。此外，槐耳有效抑制 GSH-GPX4 抗氧化系统，导致 ROS 在 PC 中大量积累。如 TEM 所示，槐耳处理的 PC 表现出线粒体嵴减少和线粒体变小，同时自噬体增加。事实上，我们发现自噬可以诱导 PC 中的铁死亡，而槐耳诱导的铁死亡可以被自噬体抑制剂 Wortmannin 抑制。

槐耳可以通过诱导 PC 中的自噬来激活铁死亡。