目的：克服肺癌细胞对凋亡和抗有丝分裂化疗的抵抗是有效治疗肺癌的关键。地菜皂甙（DG）是一种有前景的植物化学物质，可调节与肿瘤形成和进展有关的多种分子途径。然而，DG在肺癌中的精确生物学活性仍不清楚。本研究旨在调查DG在NCI-H460肺癌细胞中的抗增殖活性，探索其潜在的抗有丝分裂机制和可替代的细胞死亡途径。 在2D培养系统中，我们分析了细胞存活率、多核细胞频率、细胞浓度、细胞周期变化、细胞死亡诱导、细胞内反应性氧自由基（ROS）产生和核DNA损伤，特别是与靶基因表达相关的情况。我们还评估了DG在球体3D培养系统中的抗增殖活性，评估了体积变化、细胞死亡诱导以及抑制增殖恢复和克隆生长。 DG降低了细胞存活率和浓度，同时增加了多核细胞的频率，特别是由于子细胞融合而产生的二核细胞。这个效应与细胞负责细胞分裂的基因（RAB35、OCRL、BIRC5和AURKB）有关。此外，DG诱发的细胞死亡与坏死型凋亡（necroptosis）相关，表现为细胞内ROS产生增加以及RIPK3、MLKL、TRAF2和HSPA5基因表达的增加。在肿瘤球中，DG增加了球体体积，诱导了细胞死亡，并抑制了增殖恢复和克隆生长。 我们的研究为DG在肺癌细胞中的生物学活性提供了新的见解，有助于开发新的肿瘤学治疗方法。 版权所有 © 2023. 由Elsevier Inc.出版。

Aim Overcoming resistance to apoptosis and antimitotic chemotherapy is crucial for effective treatment of lung cancer. Diosgenin (DG), a promising phytochemical, can regulate various molecular pathways implicated in tumor formation and progression. However, the precise biological activity of DG in lung cancer remains unclear. This study aimed to investigate the antiproliferative activity of DG in NCI-H460 lung carcinoma cells to explore the underlying antimitotic mechanisms and alternative cell death pathways.In a 2D culture system, we analyzed cell viability, multinucleated cell frequency, cell concentration, cell cycle changes, cell death induction, intracellular reactive oxygen species (ROS) production, and nuclear DNA damage, particularly in relation to target gene expression. We also evaluated the antiproliferative activity of DG in a 3D culture system of spheroids, assessing volume changes, cell death induction, and inhibition of proliferation recovery and clonogenic growth.DG reduced cell viability and concentration while increasing the frequency of cells with multiple nuclei, particularly binucleated cells resulting from daughter cell fusion. This effect was associated with genes involved in cytokinesis regulation (RAB35, OCRL, BIRC5, and AURKB). Additionally, DG-induced cell death was linked to necroptosis, as evidenced by increased intracellular ROS production and RIPK3, MLKL, TRAF2, and HSPA5 gene expression. In tumor spheroids, DG increased spheroid volume, induced cell death, and inhibited proliferation recovery and clonogenic growth.Our study provides new insights into the biological activities of DG in lung cancer cells, contributing to the development of novel oncological therapies.Copyright © 2023. Published by Elsevier Inc.