葫芦素 B (CuB) 是一种在葫芦科等植物中发现的化合物，在对抗癌症（尤其是肺癌）方面表现出了良好的前景。然而，CuB 对铁死亡的具体影响及其在肺癌细胞中的作用尚未完全了解。我们的研究发现CuB可以有效减缓非小细胞肺癌（NSCLC）细胞的生长。即使少量，它也能够抑制各种非小细胞肺癌细胞系的生长。当铁死亡抑制剂 DFO、Lip-1 和 Fer-1 被引入后，这种抑制作用被逆转。 CuB 被发现可以增加 H358 肺癌细胞内活性氧 (ROS)、脂质 ROS、MDA 和亚铁离子的水平，导致 GSH、线粒体膜电位 (MMP) 降低以及铁死亡相关蛋白的变化。剂量依赖性方式。这些发现也在 A549 肺癌细胞中得到证实。在A549细胞中，不同浓度的CuB以浓度依赖性方式诱导细胞内脂质ROS、亚铁离子的积累以及铁死亡相关指标的变化。同时，CuB 在 A549 细胞中诱导的细胞毒性作用被铁死亡抑制剂 DFO 和 Fer-1 抵消。通过网络药理学，我们确定了与 CuB 处理的 NSCLC 细胞中与铁死亡相关的潜在靶点，其中 STAT3 靶点显示出高分。使用分子对接和细胞热位移测定 (CETSA) 的进一步实验表明 CuB 与 STAT3 蛋白相互作用。 Western blot 和免疫荧光染色表明 CuB 抑制 H358 细胞中 STAT3 (P-STAT3) 的磷酸化。沉默 STAT3 增强了 CuB 诱导的脂质 ROS 和铁离子的积累，以及铁死亡相关蛋白的表达。另一方面，STAT3 的过度表达逆转了 CuB 诱导的铁死亡的影响。结果表明，CuB 能够抑制 STAT3 激活，从而导致铁死亡，可能成为 NSCLC 的一种有前途的治疗选择。版权所有 © 2024。由 Elsevier B.V 出版。

Cucurbitacin B (CuB) is a compound found in plants like Cucurbitaceae that has shown promise in fighting cancer, particularly in lung cancer. However, the specific impact of CuB on ferroptosis and how it works in lung cancer cells has not been fully understood. Our research has discovered that CuB can effectively slow down the growth of non-small cell lung cancer (NSCLC) cells. Even in small amounts, it was able to inhibit the growth of various NSCLC cell lines. This inhibitory effect was reversed when ferroptosis inhibitors DFO, Lip-1 and Fer-1 were introduced. CuB was found to increase the levels of reactive oxygen species (ROS), lipid ROS, MDA, and ferrous ions within H358 lung cancer cells, leading to a decrease in GSH, mitochondrial membrane potential (MMP) and changes in ferroptosis-related proteins in a dose-dependent manner. These findings were also confirmed in A549 lung cancer cells. In A549 cells, different concentrations of CuB induced the accumulation of intracellular lipid ROS, ferrous ions and changes in ferroptosis-related indicators in a concentration-dependent manner. Meanwhile, the cytotoxic effect induced by CuB in A549 cells was counteracted by ferroptosis inhibitors DFO and Fer-1. Through network pharmacology, we identified potential targets related to ferroptosis in NSCLC cells treated with CuB, with STAT3 targets showing high scores. Further experiments using molecular docking and cell thermal shift assay (CETSA) revealed that CuB interacts with the STAT3 protein. Western blot and immunofluorescence staining demonstrated that CuB inhibits the phosphorylation of STAT3 (P-STAT3) in H358 cells. Silencing STAT3 enhanced CuB-induced accumulation of lipid ROS and iron ions, as well as the expression of ferroptosis-related proteins. On the other hand, overexpression of STAT3 reversed the effects of CuB-induced ferroptosis. The results indicate that CuB has the capability to suppress STAT3 activation, resulting in ferroptosis, and could be a promising treatment choice for NSCLC.Copyright © 2024. Published by Elsevier B.V.