Icaritin具有广泛的药理活性，包括显著的抗肿瘤活性。然而，icaritin在子宫内膜癌（UCEC）中的作用机制尚不清楚。FOX蛋白是一类高度保守的转录因子超家族，在上皮细胞分化、肿瘤转移、血管生成和细胞周期调控中起重要作用。FOXC1是FOX蛋白家族的重要成员之一。FOXC1在子宫内膜癌中异常表达，可能在子宫内膜癌的迁移和侵袭中发挥作用，然而其作用机制尚未报道。O-GlcNAc糖基化是一种常见的翻译后修饰。在子宫内膜癌中，高水平的O-GlcNAc糖基化促进细胞增殖、迁移和侵袭。癌症发展通常伴随着蛋白质的O-GlcNAc修饰，然而迄今尚未报道转录因子FOXC1的O-GlcNAc修饰。为了研究icaritin对RL95-2和Ishikawa子宫内膜癌细胞的体外和体内抑制作用，并阐明可能的分子机制，我们使用了CCK8、集落形成、迁移和侵袭实验来确定icaritin对子宫内膜癌细胞的抑制作用。细胞周期调控通过流式细胞术进行分析。蛋白水平通过西方印迹进行测量。通过免疫组化确定子宫内膜癌组织中FOXC1表达水平。为了评估icaritin是否在体内也具有活性，评估了其对肿瘤异种移植物的影响。临床样本的免疫组化分析显示，FOXC1在子宫内膜癌组织中的表达显著高于正常组织。FOXC1的下调抑制了RL95-2和Ishikawa子宫内膜癌细胞的增殖、集落形成、迁移和侵袭能力。Icaritin抑制了子宫内膜癌细胞的增殖、集落形成、迁移和侵袭，并阻断了细胞周期处于S期。Icaritin影响了FOXC1的O-GlcNAc修饰，从而影响了FOXC1的稳定性，进而抑制了子宫内膜癌细胞的增殖。Icaritin的抗子宫内膜癌作用与对FOXC1异常的O-GlcNAc修饰的抑制有关，这可能为使用icaritin对抗子宫内膜癌提供重要的理论基础。版权所有 © 2023 Elsevier GmbH发表。

Icaritin has a wide range of pharmacological activities, including significant an-titumor activity. However, the mechanism of action of icaritin in endometrial cancer (UCEC) remains unknown. FOX proteins are a highly conserved transcription factor superfamily that play important roles in epithelial cell differentiation, tumor metastasis, angiogenesis, and cell cycle regulation. FOXC1 is an important member of the FOX protein family. FOXC1 is aberrantly expressed in endometrial cancer and may play a role in the migration and invasion of endometrial cancer; however, its mechanism of action has not yet been reported. O-GlcNAc glycosylation is a common post-translational modification. In endometrial cancer, high levels of O-GlcNAcylation promote cell proliferation, migration, and invasion. Cancer development is often accompanied by O-GlcNAc modification of proteins; however, O-GlcNAc modification of the transcription factor FOXC1 has not been reported to date.To investigate the inhibitory effects of icaritin on RL95-2 and Ishikawa endometrial cancer cells in vitro and in vivo and to elucidate the possible molecular mechanisms.CCK8, colony formation, migration, and invasion assays were used to determine the inhibitory effects of icaritin on endometrial cancer cells in vitro. Cell cycle regulation was assayed by flow cytometry. Protein levels were measured based on western blotting. The level of FOXC1 expression in endometrial cancer tissues was determined by immunohistochemistry. To assess whether icaritin also has activity in vivo, its effect on tumor xenografts was evaluated.Immunohistochemical analysis of clinical samples revealed that FOXC1 expression was significantly higher in endometrial cancer tissues than in normal tissues. Downregulation of FOXC1 inhibited the proliferative, colony formation, migration, and invasive abilities of RL95-2 and Ishikawa endometrial cancer cells. Icaritin inhibited the proliferation, colony formation, migration, and invasion of endometrial cancer cells and blocked the cell cycle in S phase. Icaritin affected O-GlcNAc modification of FOXC1 and thus the stability of FOXC1, which subsequently triggered the inhibition of endometrial cancer cell proliferation.The anti-endometrial cancer effect of icaritin is related to the inhibition of abnormal O-GlcNAc modification of FOXC1, which may provide an important theoretical foundation for the use of icaritin against endometrial cancer.Copyright © 2023. Published by Elsevier GmbH.