Curcuma wenyujin Y.H. Chen & C. Ling 的根茎，在古代就被用于癌症治疗。它的化合物β-elemene (BE) 是一种有效的抗癌成分，其它化合物还包括痕量β-caryophyllene (BC)、γ-elemene 和 δ-elemene。BE 在临床治疗各种恶性肿瘤，包括肺癌时，常常与旁氏体积倍增素共同使用。EE 显示出广谱的抗癌作用，能够抑制癌细胞增殖，诱导凋亡和自噬。然而，其抗肺癌活性的确切机制尚不清楚，需要进一步的研究和探讨。本研究利用A549和PC9细胞系，通过建立裸鼠皮下肿瘤模型来评估EE在体内的疗效。利用CCK-8法确定EE及其主要成分BE和BC在不同浓度下对A549和PC9细胞的半抑制浓度（IC50）。进一步通过流式细胞术检测不同浓度的BE和BC处理的肺腺癌A549和PC9细胞的细胞周期和凋亡情况，以及线粒体膜电位（MMP）的变化。采用非靶向代谢组学分析A549细胞，探索EE及其主要活性成分BE和BC存在的潜在靶标通路和机制，然后通过试剂盒检测和Western印迹分析进行验证。本研究发现，EE、BE和BC对肺腺癌细胞的生长有明显的抑制作用，其作用机制与谷胱甘肽系统有关，通过下调与GSH合成有关的蛋白质的表达，打乱细胞氧化还原平衡，从而促进细胞凋亡。

The rhizome of Curcuma wenyujin Y.H. Chen & C. Ling, also known as Wen-E-Zhu, has been used for cancer treatment since ancient times, with roots dating back to the Song Dynasty. Elemene (EE), a sesquiterpene extract with potent anticancer properties, is extracted from Wen-E-Zhu, with β-elemene (BE) being its main active compound, along with trace amounts of β-caryophyllene (BC), γ-elemene and δ-elemene isomers. EE has demonstrated broad-spectrum anti-cancer effects and is commonly used in clinical treatments for various types of malignant cancers, including lung cancer. Studies have shown that EE can arrest the cell cycle, inhibit cancer cell proliferation, and induce apoptosis and autophagy. However, the exact mechanism of its anti-lung cancer activity remains unclear and requires further research and investigation.In this study, the possible mechanism of EE and its main active components, BE and BC, against lung adenocarcinoma was investigated by using A549 and PC9 cell lines.The subcutaneous tumor model of nude mice was constructed to evaluate the efficacy of EE in vivo, then the in vitro half-inhibitory concentration (IC50) of EE and its main active components, BE and BC, on A549 and PC9 cells at different concentrations were determined by CCK-8. Flow cytometry was used to detect the apoptosis and cycle of A549 and PC9 cells treated with different concentrations of BE and BC for 24 h. Non-targeted metabolomics analysis was performed on A549 cells to explore potential target pathways, which were subsequently verified through kit detection and western blot analysis.Injection of EE in A549 tumor-bearing mice effectively suppressed cancer growth in vivo. The IC50 of EE and its main active components, BE and BC, was around 60 μg/mL. Flow cytometry analysis showed that BE and BC blocked the G2/M and S phases of lung adenocarcinoma cells and induced apoptosis, leading to a significant reduction in mitochondrial membrane potential (MMP). Results from non-targeted metabolomics analysis indicated that the glutathione metabolism pathway in A549 cells was altered after treatment with the active components. Kit detection revealed a decrease in glutathione (GSH) levels and an increase in the levels of oxidized glutathione (GSSG) and reactive oxygen (ROS). Supplementation of GSH reduced the inhibitory activity of the active components on lung cancer and also decreased the ROS content of cells. Analysis of glutathione synthesis-related proteins showed a decrease in the expression of glutaminase, cystine/glutamate reverse transporter (SLC7A11), and glutathione synthase (GS), while the expression of glutamate cysteine ligase modified subunit (GCLM) was increased. In the apoptosis-related pathway, Bax protein and cleaved caspase-9/caspase-9 ratio were up-regulated and Bcl-2 protein was down-regulated.EE, BE, and BC showed significant inhibitory effects on the growth of lung adenocarcinoma cells, and the mechanism of action was linked to the glutathione system. By down-regulating the expression of proteins related to GSH synthesis, EE and its main active components BE and BC disrupted the cellular redox system and thereby promoted cell apoptosis.Copyright © 2023. Published by Elsevier B.V.