Silencing of FZD7 Inhibits Endometriotic Cell Viability, Migration, and Angiogenesis by Promoting Ferroptosis.

IF 1.8 4区生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Cell Biochemistry and Biophysics Pub Date : 2025-01-11 DOI:10.1007/s12013-024-01656-4

Yi Zhang, Huifen Yang

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Abstract

Background: 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.

Methods: 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 Fe²⁺ 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).

Results: 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 Fe²⁺ 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.

Conclusion: The above indices suggest that FZD7 knockdown regulates endometriotic cell proliferation, invasion, migration, and angiogenesis via ferroptosis.

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沉默FZD7通过促进铁下垂抑制子宫内膜异位症细胞活力、迁移和血管生成。

背景：子宫内膜异位症（EMS）是一种较难治愈的妇科疾病。卷曲-7 （FZD7）已被证明与EMS的发展有关，但其具体机制尚不清楚。本研究旨在探讨FZD7在EMS中的作用。方法：采用RT-qPCR和western blot检测FZD7在人子宫内膜基质细胞（hESCs）和人异位子宫内膜基质细胞系hEM15A中的表达水平。建立了FZD7的干扰质粒。采用CCK-8、EdU、伤口愈合、跨井侵袭和细胞骨架染色法评估FZD7沉默在hEM15A细胞增殖、侵袭和迁移中的作用。用成管实验评价细胞成管能力。采用试剂盒检测细胞VEGF、GSH、MDA水平。分别用C11-BODIPY（581/591）和FeRhoNox-1探针检测细胞内脂质ROS和Fe2+水平。western blot检测凋亡相关蛋白SLC7A11、GPX4和ACSL4的表达。通过添加铁下垂抑制剂（fer1）进一步分析铁下垂对子宫内膜异位症细胞活力、迁移和血管生成的影响。结果：FZD7在hEM15A细胞中表达上调，沉默FZD7可抑制细胞增殖、迁移、侵袭和血管生成能力。下调FZD7可降低细胞GSH水平，升高MDA水平。FZD7的下调还导致细胞内ROS和Fe2+水平升高，SLC7A11和GPX4水平下调，ACSL4水平上调，这些都是铁下垂的标志。然而，当加入铁下垂抑制剂fe -1时，FZD7敲低对hEM15A细胞进展的抑制作用被逆转。结论：FZD7敲低可通过铁下垂调节子宫内膜异位症细胞的增殖、侵袭、迁移和血管生成。

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来源期刊

Cell Biochemistry and Biophysics 生物-生化与分子生物学

CiteScore

4.40

自引率

0.00%

发文量

审稿时长

7.5 months

期刊介绍： Cell Biochemistry and Biophysics (CBB) aims to publish papers on the nature of the biochemical and biophysical mechanisms underlying the structure, control and function of cellular systems The reports should be within the framework of modern biochemistry and chemistry, biophysics and cell physiology, physics and engineering, molecular and structural biology. The relationship between molecular structure and function under investigation is emphasized. Examples of subject areas that CBB publishes are: · biochemical and biophysical aspects of cell structure and function; · interactions of cells and their molecular/macromolecular constituents; · innovative developments in genetic and biomolecular engineering; · computer-based analysis of tissues, cells, cell networks, organelles, and molecular/macromolecular assemblies; · photometric, spectroscopic, microscopic, mechanical, and electrical methodologies/techniques in analytical cytology, cytometry and innovative instrument design For articles that focus on computational aspects, authors should be clear about which docking and molecular dynamics algorithms or software packages are being used as well as details on the system parameterization, simulations conditions etc. In addition, docking calculations (virtual screening, QSAR, etc.) should be validated either by experimental studies or one or more reliable theoretical cross-validation methods.