背景:子宫内膜癌(EC)是女性生殖系统中最常见的恶性肿瘤之一,通常有一个糟糕的诊断和不利的健康影响。Neferine(Nef),来源于食用和药用莲子,以其功能活性而闻名;然而,其对EC的抗癌机制仍然难以捉摸。
目的:我们探索了Nef对EC的潜在抗癌作用和潜在的分子机制。
方法:使用MTT测试细胞毒性,和细胞周期,凋亡,Ca2+水平,通过流式细胞术观察线粒体膜电位(MMP)。Nef治疗后,使用miRNA-seq数据鉴定miRNA表达的差异.此外,免疫印迹和免疫组织化学(IHC)用于鉴定与小鼠和细胞凋亡相关的蛋白。
结果:Nef处理导致Ishikawa细胞凋亡并阻断G2/M期细胞增殖。总的来说,获得101个显著不同的miRNA(p〈0.05和|logFC|〉1),并进行GO和KEGG富集分析,揭示了与细胞凋亡有关的Ca2+和PI3K/AKT信号通路。Nef处理显著改变细胞内Ca2+和MMP,激活内质网应激(ERS)通路和线粒体通路中关键蛋白的表达。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的表达式,Bcl-2,Caspase3,Cyto-c,和p-AKT也与体外结果一致。
结论:Nef可阻断细胞周期并诱导线粒体凋亡途径的激活,涉及Ca2+介导的ERS途径和PI3K/AKT途径,从而诱导EC细胞凋亡,确认Nef在预防和治疗EC中的潜在作用。
BACKGROUND: 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.
OBJECTIVE: We explored the potential anti-cancer effects and underlying molecular mechanisms of Nef on EC.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: 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.