Zhang Yi, Yang Huifen
Department of Obstetrics and Gynecology, The Affiliated Women and Children's Hospital of Ningbo University, Ningbo, Zhejiang, 315012, China.
Department of Obstetrics and Gynecology, Lishui Municipal Central Hospital, Lishui, Zhejiang, 323000, China.
Cell Biochem Biophys. 2025 Jun;83(2):2471-2480. doi: 10.1007/s12013-024-01656-4. Epub 2025 Jan 11.
Endometriosis (EMS) is a difficult gynecological disease to cure. Frizzled-7 (FZD7) has been shown to be associated with the development of EMS, but its specific mechanism remains unclarified. This study aims to explore the role of FZD7 in EMS.
RT-qPCR and western blot were used to detect the expression level of FZD7 in human endometrial stromal cells (hESCs) and human ectopic endometrial stromal cell line hEM15A. The interfering plasmid of FZD7 was established. CCK-8, EdU, wound healing, transwell invasion, and cytoskeletal staining assays were applied to evaluate the role of FZD7 silencing in hEM15A cell proliferation, invasion, and migration. Tube forming ability of cells was evaluated by tube formation assay. Cellular VEGF, GSH, and MDA levels were measure by kits. Intracellular lipid ROS and Fe levels were tested using C11-BODIPY (581/591) and FeRhoNox-1 probes, respectively. The ferroptosis-related protein SLC7A11, GPX4, and ACSL4 expressions were analyzed using western blot. The effects of ferroptosis on endometriotic cell viability, migration, and angiogenesis were further analyzed with the addition of an ferroptosis inhibitor (Fer-1).
FZD7 was upregulated in hEM15A cells, and silencing of FZD7 inhibited cell proliferation, migration, invasion, and angiogenesis abilities. Downregulation of FZD7 decreased cellular GSH level and elevated MDA level. Knockdown of FZD7 also caused an increase in intracellular ROS and Fe levels, as well as the downregulation of SLC7A11 and GPX4 levels and the upregulation of ACSL4 level, which are hallmarks of ferroptosis. However, the inhibitory effects of FZD7 knockdown on hEM15A cell progression were reversed when ferroptosis inhibitor Fer-1 added.
The above indices suggest that FZD7 knockdown regulates endometriotic cell proliferation, invasion, migration, and angiogenesis via ferroptosis.
子宫内膜异位症（EMS）是一种难以治愈的妇科疾病。卷曲蛋白7（FZD7）已被证明与EMS的发生发展有关，但其具体机制仍不清楚。本研究旨在探讨FZD7在EMS中的作用。
采用RT-qPCR和蛋白质免疫印迹法检测人子宫内膜基质细胞（hESCs）和人异位子宫内膜基质细胞系hEM15A中FZD7的表达水平。构建FZD7干扰质粒。应用CCK-8、EdU、伤口愈合、Transwell侵袭和细胞骨架染色试验评估FZD7沉默对hEM15A细胞增殖、侵袭和迁移的作用。通过管腔形成试验评估细胞的管腔形成能力。使用试剂盒检测细胞VEGF、GSH和MDA水平。分别使用C11-BODIPY（581/591）和FeRhoNox-1探针检测细胞内脂质活性氧和铁水平。采用蛋白质免疫印迹法分析铁死亡相关蛋白SLC7A11、GPX4和ACSL4的表达。添加铁死亡抑制剂（Fer-1）进一步分析铁死亡对子宫内膜异位症细胞活力、迁移和血管生成的影响。
FZD7在hEM15A细胞中上调，FZD7沉默抑制细胞增殖、迁移、侵袭和血管生成能力。FZD7下调降低细胞GSH水平，升高MDA水平。FZD7敲低还导致细胞内活性氧和铁水平升高，以及SLC7A11和GPX4水平下调和ACSL4水平上调，这些都是铁死亡的标志。然而，添加铁死亡抑制剂Fer-1后，FZD7敲低对hEM15A细胞进程的抑制作用被逆转。
上述指标表明，FZD7敲低通过铁死亡调节子宫内膜异位症细胞的增殖、侵袭、迁移和血管生成。
