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

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. Similar content being viewed by others Data Availability The data in the present study are available from the corresponding author on reasonable request.

As-Sanie, S., et al. (2019). Assessing research gaps and unmet needs in endometriosis. American Journal of Obstetrics & Gynecology, 221(2), 86–94. Somigliana, E., et al. (2006). Association between endometriosis and cancer: a comprehensive review and a critical analysis of clinical and epidemiological evidence. Gynecologic Oncology, 101(2), 331–341. Liu, H., et al. (2019). GLI1 is increased in ovarian endometriosis and regulates migration, invasion and proliferation of human endometrial stromal cells in endometriosis. Annals of Translational Medicine, 7(22), 663. Marí-Alexandre, J., et al. (2015). MicroRNAs and angiogenesis in endometriosis. Thrombosis Research, 135(Suppl 1), S38–S40. Vercellini, P., et al. (2014). Endometriosis: pathogenesis and treatment. Nature Review Endocrinology, 10(5), 261–275. Pirtea, P., et al. (2022). Infertility workup: identifying endometriosis. Fertility and Sterility, 118(1), 29–33. Amro, B., et al. (2022). New Understanding of Diagnosis, Treatment and Prevention of Endometriosis. International of J Environmental Research Public Health, 19(11), 6725. Mechsner, S. (2022). Endometriosis, an Ongoing Pain-Step-by-Step Treatment. Journal of Clinical Medicine, 11(2), 467. Stockwell, B. R., et al. (2017). Ferroptosis: A Regulated Cell Death Nexus Linking Metabolism, Redox Biology, and Disease. Cell, 171(2), 273–285. Rochette, L., et al. (2022). Lipid Peroxidation and Iron Metabolism: Two Corner Stones in the Homeostasis Control of Ferroptosis. International Journal of Molecular Science, 24(1), 449. Defrère, S., et al. (2008). Potential involvement of iron in the pathogenesis of peritoneal endometriosis. Molecular Human Reproduction, 14(7), 377–385. Li, Y., et al. (2023). Double-edged roles of ferroptosis in endometriosis and endometriosis-related infertility. Cell Death Discovery, 9(1), 306. Lousse, J. C., et al. (2009). Iron storage is significantly increased in peritoneal macrophages of endometriosis patients and correlates with iron overload in peritoneal fluid. Fertility and Sterility, 91(5), 1668–1675. Ng, S. W., et al. (2020). Endometriosis: The Role of Iron Overload and Ferroptosis. Reproductive Science, 27(7), 1383–1390. Burney, R. O., & Giudice, L. C. (2012). Pathogenesis and pathophysiology of endometriosis. Fertility and Sterility, 98(3), 511–519. Zou, W., et al. (2024). Resveratrol protected against the development of endometriosis by promoting ferroptosis through miR-21-3p/p53/SLC7A11 signaling pathway. Biochemical and Biophysical Research Communication, 692, 149338. Lan, S., Zhang, Z., & Li, Q. (2023). FZD7: A potential biomarker for endometriosis. Medicine (Baltimore), 102(40), e35406. Bae, S. J., et al. (2022). Identification and analysis of novel endometriosis biomarkers via integrative bioinformatics. Frontiers in Endocrinology (Lausanne), 13, 942368. Zhu, H., et al. (2019). MicroRNA-488 inhibits endometrial glandular epithelial cell proliferation, migration, and invasion in endometriosis mice via Wnt by inhibiting FZD7. Journal of Cellular and Molecular Medicine, 23(4), 2419–2430. Guo, S., et al. (2023). The malignancy suppression and ferroptosis facilitation of BCL6 in gastric cancer mediated by FZD7 repression are strengthened by RNF180/RhoC pathway. Cell & Bioscience, 13(1), 73. Li, J., et al. (2021). Tumor heterogeneity in autophagy-dependent ferroptosis. Autophagy, 17(11), 3361–3374. Chapron, C., et al. (2019). Rethinking mechanisms, diagnosis and management of endometriosis. Nature Review Endocrinology, 15(11), 666–682. Asad, M., et al. (2014). FZD7 drives in vitro aggressiveness in Stem-A subtype of ovarian cancer via regulation of non-canonical Wnt/PCP pathway. Cell Death Disease, 5(7), e1346. Yang, L., et al. (2011). FZD7 has a critical role in cell proliferation in triple negative breast cancer. Oncogene, 30(43), 4437–4446. Peghaire, C., et al. (2016). Fzd7 (Frizzled-7) Expressed by Endothelial Cells Controls Blood Vessel Formation Through Wnt/β-Catenin Canonical Signaling. Arteriosclerosis Thrombosis & Vascular Biology, 36(12), 2369–2380. Xie, W., et al. (2021). A novel humanized Frizzled-7-targeting antibody enhances antitumor effects of Bevacizumab against triple-negative breast cancer via blocking Wnt/β-catenin signaling pathway. Journal of Experimental & Clinical Cancer Research, 40(1), 30. Li, B., et al. (2021). Ferroptosis resistance mechanisms in endometriosis for diagnostic model establishment. Reproductive Biomedicine Online, 43(1), 127–138. Zhang, W., et al. (2021). FOXM1D potentiates PKM2-mediated tumor glycolysis and angiogenesis. Molecular Oncology, 15(5), 1466–1485. Wan, Y., et al. (2022). Long noncoding RNA ADAMTS9-AS1 represses ferroptosis of endometrial stromal cells by regulating the miR-6516-5p/GPX4 axis in endometriosis. Science Reports, 12(1), 2618. Zhang, D., et al. (2022). MiR-1-3p enhances the sensitivity of ovarian cancer cells to ferroptosis by targeting FZD7. Zhong Nan Da Xue Xue Bao Yi Xue Ban, 47(11), 1512–1521. Funding This work was supported by The Zhejiang Provincial Medical and Health Science and Technology Plan Project (No: 2025KY1938). Author information Authors and Affiliations Contributions Yi Zhang designed the study and analyzed data Huifen Yang graphing and wrote the manuscript. All authors reviewed and revised the manuscript, and approved the final manuscript. Corresponding author Ethics declarations Conflict of interest The authors declare no competing interests. Additional information Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations. Rights and permissions Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law. About this article Cite this article Zhang, Y., Yang, H. Silencing of FZD7 Inhibits Endometriotic Cell Viability, Migration, and Angiogenesis by Promoting Ferroptosis. Cell Biochem Biophys 83, 2471–2480 (2025). https://doi.org/10.1007/s12013-024-01656-4 Accepted: Published: Version of record: Issue date: DOI: https://doi.org/10.1007/s12013-024-01656-4