胃癌是一种侵袭性恶性肿瘤，是癌症死亡的主要原因。 Ephrin-A4（EFNA4）被认为与胃癌的免疫微环境和预后有关。本研究旨在探讨EFNA4在胃癌发生发展中的参与及其机制。 RT-qPCR 和蛋白质印迹检测了胃癌细胞中 EFNA4 和 Pygopu​​s2 (Pygo2) 的表达。转染EFNA4干扰质粒或共转染EFNA4干扰质粒和Pygo2过表达质粒后，采用CCK-8法和EDU染色检测细胞增殖情况。伤口愈合、Transwell、TUNEL 和内皮细胞管形成实验分别检测细胞迁移、侵袭、凋亡和血管生成。蛋白质印迹检查了转移、凋亡、血管生成和 Wnt 信号相关蛋白的表达。通过球体形成测定、RT-qPCR 和蛋白质印迹评估细胞干性。通过实验数据，注意到胃癌细胞中EFNA4表达增加。 EFNA4的敲低抑制了胃癌细胞的增殖、迁移、侵袭、血管生成以及干性，同时加剧了细胞凋亡。此外，EFNA4 缺失会降低 Pygo2 蛋白表达，然后使 Wnt/β-catenin 信号转导失活。 Pygo2 的进一步升高逆转了 EFNA4 沉默对 Wnt/β-catenin 信号传导、细胞增殖、凋亡、迁移、侵袭、血管生成以及胃癌干性的影响。因此，敲低 EFNA4 可能会下调 Pygo2 并使 Wnt/β-catenin 信号失活，从而发挥针对胃癌的保护作用。©作者 2024。开放获取。本文根据知识共享 CC-BY 国际许可证获得许可。

Gastric cancer represents an aggressive malignancy and a leading contributor to cancer death. Ephrin-A4 (EFNA4) has been proposed to be related to the immune microenvironment and prognosis of gastric cancer. This study was undertaken to discuss the participation and mechanism of EFNA4 in the development of gastric cancer. RT-qPCR and western blot examined EFNA4 and Pygopus2 (Pygo2) expression in gastric cancer cells. After transfection of EFNA4 interference plasmids or co-transfection of EFNA4 interference plasmids and Pygo2 overexpression plasmids, cell proliferation was detected by the CCK-8 method and EDU staining. Wound healing, Transwell, TUNEL, and endothelial cell tube formation assays detected cell migration, invasion, apoptosis, and angiogenesis, respectively. Western blot examined the expression of metastasis-, apoptosis-, angiogenesis-, and Wnt signaling-associated proteins. Cell stemness was estimated by the sphere formation assay, RT-qPCR, and western blot. Through the experimental data, it was noticed that EFNA4 expression was increased in gastric cancer cells. Knockdown of EFNA4 suppressed the proliferation, migration, invasion, angiogenesis as well as stemness while aggravating the apoptosis of gastric cancer cells. Also, EFNA4 depletion reduced Pygo2 protein expression and then inactivated Wnt/β-catenin signaling. Further elevation of Pygo2 reversed the impacts of EFNA4 silencing on Wnt/β-catenin signaling, cell proliferation, apoptosis, migration, invasion, angiogenesis as well as stemness in gastric cancer. Accordingly, the knockdown of EFNA4 might downregulate Pygo2 and inactivate Wnt/β-catenin signaling to exert protective effects against gastric cancer.©The Author(s) 2024. Open Access. This article is licensed under a Creative Commons CC-BY International License.