萋芳酮（ZER）是从姜科植物中分离出来的植物化学物质。许多研究表明它具有多样的药理学特性，特别是强大的抗肿瘤活性。本研究旨在评估ZER对HT-29细胞在二维（2D）单层和三维（3D）球状培养系统中的抗增殖效应。通过对3D球体HT-29细胞的生长（大小）、细胞死亡和细胞周期阻滞的评估与mRNA表达数据相关联。对2D细胞的处理显示，ZER在30 µM以上的浓度下表现出细胞毒性，并且83.54 µM的半数抑制浓度（ZER处理后24小时）有效抑制了细胞迁移。在3D模型中，ZER导致了72小时内球体体积的增加，这归因于特征区域的解聚和重组。细胞死亡分析显示，在ZER处理后24小时，凋亡细胞显著增加，并且细胞周期在G1期阻滞。此外，ZER处理导致mRNA表达的改变，影响了涉及细胞死亡（BCL2和BBC3）、内质网应激（ERN1）、DNA损伤（GADD45A）、细胞周期调控（CDKN1A、NFKB1、MYC和TP53）和自噬（BECN1和SQSTM1）的关键信号通路。这些发现表明，ZER有望成为开发新型抗癌药物的潜在候选物，可以调节重要的细胞信号通路。此外，使用3D培养系统在我们的研究中证明是一种有价值的工具。© 2023年. 作者（或作者）独家授权给施普林格-弗朗西斯出版公司德国部分，施普林格出版公司的一部分。

Zerumbone (ZER) is a phytochemical isolated from plants of the Zingiberaceae family. Numerous studies have demonstrated its diverse pharmacological properties, particularly its potent antitumorigenic activity. This study aimed to assess the antiproliferative effects of ZER on HT-29 cells cultivated in both two-dimensional (2D) monolayer and three-dimensional (3D) spheroid culture systems. The evaluation of growth (size), cell death, and cell cycle arrest in 3D spheroid HT-29 cells was correlated with mRNA expression data. Treatment of 2D cells revealed that ZER exhibited cytotoxicity at concentrations above 30 µM, and an IC50 of 83.54 µM (24-h post-ZER treatment) effectively suppressed cell migration. In the 3D model, ZER induced an increase in spheroid volume over a 72-h period attributed to disaggregation and reconfiguration of characteristic zones. Analysis of cell death demonstrated a significant rise in apoptotic cells after 24 h of ZER treatment, along with cell cycle arrest in the G1 phase. Furthermore, ZER treatment resulted in alterations in mRNA expression, affecting key signaling pathways involved in cell death (BCL2 and BBC3), endoplasmic reticulum stress (ERN1), DNA damage (GADD45A), cell cycle regulation (CDKN1A, NFKB1, MYC, and TP53), and autophagy (BECN1 and SQSTM1). These findings suggested that ZER holds promise as a potential candidate for the development of novel anticancer agents that can modulate crucial cell signaling pathways. Additionally, the use of the 3D culture system proved to be a valuable tool in our investigation.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.