Department of Obstetrics and Gynecology, Jinshan Hospital of Fudan University, Shanghai 201508, People's Republic of China.
Department of Reproductive Immunology, The International Peace Maternity and Child Health Hospital, School of Medicine, Shanghai Jiao Tong University, Shanghai 200030, People's Republic of China.
J Reprod Immunol. 2024 Sep;165:104312. doi: 10.1016/j.jri.2024.104312. Epub 2024 Jul 24.
Endometriosis (EMs) is a chronic disease characterized by endometrial-like tissue present outside of the uterus. Macrophages have been confirmed to participate in the development of EMs. Integrin β3 (ITGB3), a β-subunit of the integrin family, is crucial in tumor progression. In this study, we investigated the pivotal role of ITGB3 in endometrial stromal cells (ESCs) and its influence on the development of EMs, particularly focusing on the regulatory impact of macrophages.
In this study, we used western blot, Real-time qPCR, Immunohistochemistry to detected the high expression of ITGB3 in ESCs. ITGB3-overexpression ESCs (ITGB3-OE) was constructed and detected by RNA-seq with normal ESCs. ATP and lactate expression assay, transwell migration assay, wound healing, cell adhesion assay and other molecular biology techniques were used to explore the potential mechanisms. In vivo, we constructed the EMs mouse model and injected with cilengitite to inhibit ITGB3.
Here, we found ITGB3 highly expressed in ectopic lesions in EMs. The increasing ITGB3 resulted in activating the glycolysis, which produced more ATP and lactate in ITGB3-OE. After culturing with lactate, the migration, proliferation and invasion ability of ESCs were enhanced, while the result in 2-DG was reversed. In vivo, the results showed that after antagonizing ITGB3, the number of ectopic lesions was decrease.
Our findings indicate that ITGB3 up-regulated by macrophages are able to regulate the glycolysis to promote the development of EMs and lactate enhances the ability of proliferation, migration, invasion and adhesion of EMs iv vivo and in vitro.
子宫内膜异位症(EMs)是一种以子宫内膜样组织出现在子宫外为特征的慢性疾病。已证实巨噬细胞参与了 EMs 的发展。整合素 β3(ITGB3)是整合素家族的β亚基,在肿瘤进展中至关重要。在这项研究中,我们研究了 ITGB3 在子宫内膜基质细胞(ESCs)中的关键作用及其对 EMs 发展的影响,特别是聚焦于巨噬细胞的调节作用。
在这项研究中,我们使用 Western blot、实时 qPCR 和免疫组织化学检测到 ESCs 中 ITGB3 的高表达。通过 RNA-seq 构建并检测 ITGB3 过表达 ESCs(ITGB3-OE)与正常 ESCs。通过 ATP 和乳酸表达测定、Transwell 迁移测定、划痕愈合测定、细胞黏附测定和其他分子生物学技术,我们探讨了潜在的机制。在体内,我们构建了 EMs 小鼠模型,并注射 cilengitide 抑制 ITGB3。
在这里,我们发现 ITGB3 在 EMs 的异位病变中高表达。增加的 ITGB3 导致糖酵解激活,从而在 ITGB3-OE 中产生更多的 ATP 和乳酸。在与乳酸共培养后,ESCs 的迁移、增殖和侵袭能力增强,而 2-DG 的结果则相反。在体内,结果表明拮抗 ITGB3 后,异位病变的数量减少。
我们的研究结果表明,巨噬细胞上调的 ITGB3 能够调节糖酵解以促进 EMs 的发展,而乳酸增强了 EMs 在体内和体外的增殖、迁移、侵袭和黏附能力。