Cancer Institute of Traditional Chinese Medicine, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Department of Surgery, University of California, La Jolla, San Diego, CA, United States; AntiCancer Inc., San Diego, CA, United States.
Phytomedicine. 2023 Jan;108:154493. doi: 10.1016/j.phymed.2022.154493. Epub 2022 Oct 10.
Esophageal squamous cell carcinoma (ESCC) is a malignancy with high incidence in several regions of China, and the prognosis of patients with ESCC is unfavorable. Evodiamine (Evo), a small molecule derived from the traditional Chinese herb Evodia rutaecarpa, has shown anti-cancer efficacy in numerous human malignancies but not in ESCC.
To determine whether Evo induces cell-cycle arrest and apoptosis in ESCC in vitro and in vivo and elucidate the underlying mechanisms.
ATPlite and colony formation assay were used to validate the inhibiting effect of Evo on three ESCC cells in vitro; Two subcutaneous tumor models of ESCC cells were used to evaluate the anti-ESCC effect of Evo and assess the biosafety of Evo in vivo; RNAseq and Database of KEGG pathway analysis provided a direction for the mechanistic study of Evo; FACS was used to detect Evo-induced cell cycle arrest and cell apoptosis in ESCC cells; Western blot and QPCR were respectively used to detect the level of related genes and proteins in Evo-treated ESCC cells; SiRNA and other experimental techniques were used to identify the molecular mechanism of Evo-induced ESCC cell cycle arrest and cell apoptosis.
Evo significantly suppressed the growth of ESCC both in vitro and in vivo. Mechanistically, Evo induced M-phase cell-cycle arrest by inactivation of CUL4A E3 ligase, which mediates degradation blockage of p53 and transcriptional activation of p21. With the prolonged treatment time, Evo triggered both Noxa-dependent intrinsic and DR4-dependent extrinsic cell apoptosis in two ESCC cell lines.
Our findings revealed the anti-tumor efficacy and mechanisms of Evo, providing a solid scientific basis for Evo as an attractive choice for ESCC treatment.
食管鳞状细胞癌（ESCC）是中国多个地区发病率较高的恶性肿瘤，ESCC 患者的预后不佳。吴茱萸碱（Evo）是一种源自传统中药吴茱萸的小分子，已在多种人类恶性肿瘤中显示出抗癌疗效，但在 ESCC 中没有。
确定 Evo 是否在体外和体内诱导 ESCC 细胞周期停滞和细胞凋亡，并阐明其潜在机制。
使用 ATPlite 和集落形成测定法验证 Evo 对三种 ESCC 细胞的抑制作用；使用两种 ESCC 细胞的皮下肿瘤模型评估 Evo 的抗 ESCC 作用，并评估 Evo 在体内的生物安全性；RNAseq 和 KEGG 途径分析数据库为 Evo 的机制研究提供了方向；FACS 用于检测 Evo 诱导的 ESCC 细胞周期停滞和细胞凋亡；Western blot 和 QPCR 分别用于检测 Evo 处理的 ESCC 细胞中相关基因和蛋白的水平；SiRNA 等实验技术用于鉴定 Evo 诱导的 ESCC 细胞周期停滞和细胞凋亡的分子机制。
Evo 显著抑制了 ESCC 的体外和体内生长。在机制上，Evo 通过失活 CUL4A E3 连接酶诱导 M 期细胞周期停滞，该酶介导 p53 的降解阻断和 p21 的转录激活。随着治疗时间的延长，Evo 在两种 ESCC 细胞系中引发了依赖 Noxa 的内在和依赖 DR4 的外在细胞凋亡。
我们的研究结果揭示了 Evo 的抗肿瘤功效和机制，为 Evo 作为 ESCC 治疗的一种有吸引力的选择提供了坚实的科学基础。
