子宫内膜癌（EC）作为女性生殖系统中最常见的恶性肿瘤之一，通常诊断不佳并对健康产生不利影响。 Neferine (Nef) 源自食用和药用莲子，因其功能活性而闻名；然而，其对 EC 的抗癌机制仍不明确。我们探讨了 Nef 对 EC 的潜在抗癌作用及其潜在分子机制。采用 MTT 测试细胞毒性，并通过细胞周期、细胞凋亡、Ca2 水平和线粒体膜进行检测。通过流式细胞仪观察MMP电位。 Nef 处理后，使用 miRNA-seq 数据鉴定了 miRNA 表达的差异。此外，采用蛋白质印迹和免疫组织化学（IHC）来鉴定与小鼠和细胞凋亡相关的蛋白质。Nef处理导致Ishikawa细胞凋亡并阻断G2/M期的细胞增殖。总共获得了101个显着差异的miRNA（p<0.05和|logFC|>1），并进行GO和KEGG富集分析，揭示了与细胞凋亡相关的Ca2和PI3K/AKT信号通路。 Nef 处理显着改变细胞内 Ca2+ 水平和 MMP，激活内质网应激 (ERS) 途径和线粒体途径中关键蛋白的表达。此外，Nef还抑制PI3K/AKT通路中关键蛋白的表达，引起细胞凋亡。此外，在小鼠肿瘤组织中，CHOP、Bcl-2、Caspase 3、Cyto-c、p-AKT的表达也与体外结果一致。Nef可以阻断细胞周期，诱导线粒体凋亡的激活该通路涉及 Ca2 介导的 ERS ​​通路和 PI3K/AKT 通路，从而诱导 EC 细胞凋亡，证实了 Nef 在预防和治疗 EC 中的潜在作用。版权所有 © 2024 作者。由 Elsevier GmbH 出版。保留所有权利。

Endometrial cancer (EC) as one of the most prevalent malignancies in the female reproductive system, usually has a poor diagnosis and unfavorable health effects. Neferine (Nef), derived from the edible and medicinal lotus seed, has been known for its functional activity; however, its anti-cancer mechanism for EC remains elusive.We explored the potential anti-cancer effects and underlying molecular mechanisms of Nef on EC.The cytotoxicity was tested using MTT, and the cell cycle, apoptosis, Ca2+ levels, and the mitochondrial membrane potential (MMP) were observed through flow cytometry. After Nef treatment, differences in miRNA expression were identified using miRNA-seq data. Furthermore, western blot and immunohistochemistry (IHC) were employed to identify the proteins associated with apoptosis in both mice and cells.Nef treatment led to Ishikawa cell apoptosis and blocked cell proliferation in the G2/M phase. In total, 101 significantly different miRNA (p 〈 0.05 and |logFC| 〉 1) were obtained and subjected to GO and KEGG enrichment analysis, which revealed the Ca2+ and PI3K/AKT signaling pathways pertaining to apoptosis. Nef treatment significantly changed intracellular Ca2+ levels and MMP, activating the endoplasmic reticulum stress (ERS) pathway and the expression of key proteins in the mitochondrial pathway. In addition, Nef also inhibited the expression of key proteins in the PI3K/AKT pathway, causing cell apoptosis. Moreover, in mouse tumor tissues, the expression of CHOP, Bcl-2, Caspase 3, Cyto-c, and p-AKT was also consistent with the results in vitro.Nef could block the cell cycle and induce the activation of the mitochondrial apoptotic pathway involving the Ca2+-mediated ERS pathway and the PI3K/AKT pathway, thereby inducing apoptosis in EC cells, confirming the potential role of Nef in the prevention and treatment of EC.Copyright © 2024 The Author(s). Published by Elsevier GmbH.. All rights reserved.