Departments of Prenatal Diagnosis, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.
Departments of Laboratory Diagnosis, The Second Affiliated Hospital of Harbin Medical University, Harbin, Heilongjiang 150001, P.R. China.
Mol Med Rep. 2018 Jun;17(6):7505-7512. doi: 10.3892/mmr.2018.8822. Epub 2018 Mar 29.
Antiphospholipid antibody (aPL)‑mediated antiphospholipid syndrome (APS) is an autoimmune disease. Upon binding to aPL, the primary antigen of aPL, β2‑glycoprotein I (β2‑GP I), induces abnormal immune function, which further activates downstream signaling pathways in the cell and eventually leads to APS. The present study aimed to determine whether β2‑GP I antigen and anti‑β2‑glycoprotein I antibody (aβ2‑GP I), which belong to the aPL class of antibodies, may affect human chorionic epithelium cell (JEG‑3) proliferation, migration and invasion. Recombinant human (rh)β2‑GP I protein was expressed using a prokaryotic expression system and aβ2‑GP I antibody was purified from the blood serum of 10 patients with recurrent pregnancy loss. JEG‑3 cells were stimulated with rhβ2‑GP I and aβ2‑GP I separately or simultaneously, and serum immunoglobulin G of normal pregnant women was used as negative control. Using cell counting kit‑8, cell cycle and transwell assays in addition to EdU staining, it was determined that aβ2‑GP I/rhβ2‑GP I complex markedly increased JEG‑3 cell proliferation, migration and invasion. The results revealed that mRNA levels of inhibitor of nuclear factor (NF)‑κB kinase subunit (IKKβ), myeloid differentiation primary response protein MyD88 (MyD88), NF‑κB and NF‑κB inhibitor α (IκBα), as well as the protein levels of MyD88, IκBα and phospho(p)‑IκBα in JEG‑3 cells increased following incubation with the aβ2‑GP I/rhβ2‑GP I complex. The observed upregulation of p‑IκBα protein suggested that IκBα‑mediated inhibition of NF‑κB was weakened. Furthermore, JEG‑3 cells were transfected with PGMLV‑NF‑κB‑Lu vector. Luciferase activity in JEG‑3‑NFκB‑Luc1 and JEG‑3‑NFκB‑Luc2 cells was enhanced following treatment with aβ2‑GP I/rhβ2‑GP I complex. The present study demonstrated that aβ2‑GP I/rhβ2‑GP I complex activates NF‑κB through MyD88 signal transduction pathway, which further enhances JEG‑3 cell proliferation, migration and invasion.
抗磷脂抗体(aPL)介导的抗磷脂综合征(APS)是一种自身免疫性疾病。当与 aPL 的主要抗原β2-糖蛋白 I(β2-GPI)结合时,aPL 会诱导异常的免疫功能,从而进一步激活细胞内的下游信号通路,最终导致 APS。本研究旨在确定β2-GPI 抗原和抗β2-糖蛋白 I 抗体(aβ2-GPI)是否会影响人绒毛膜上皮细胞(JEG-3)的增殖、迁移和侵袭。使用原核表达系统表达重组人(rh)β2-GPI 蛋白,并从 10 例复发性流产患者的血清中纯化 aβ2-GPI 抗体。分别用 rhβ2-GPI 和 aβ2-GPI 刺激 JEG-3 细胞,并用正常孕妇血清作为阴性对照。通过细胞计数试剂盒-8、细胞周期和 Transwell 检测以及 EdU 染色,确定 aβ2-GPI/rhβ2-GPI 复合物显著增加了 JEG-3 细胞的增殖、迁移和侵袭。结果表明,rhβ2-GPI/aβ2-GPI 复合物孵育后,JEG-3 细胞中核因子(NF)-κB 激酶亚单位(IKKβ)、髓样分化初级反应蛋白 MyD88(MyD88)、NF-κB 和 NF-κB 抑制剂α(IκBα)的 mRNA 水平以及 MyD88、IκBα 和磷酸化(p)-IκBα 的蛋白水平均升高。观察到 p-IκBα 蛋白的上调表明 IκBα 介导的 NF-κB 抑制减弱。此外,用 PGMLV-NF-κB-Lu 载体转染 JEG-3 细胞。用 aβ2-GPI/rhβ2-GPI 复合物处理后,JEG-3-NFκB-Luc1 和 JEG-3-NFκB-Luc2 细胞中的荧光素酶活性增强。本研究表明,aβ2-GPI/rhβ2-GPI 复合物通过 MyD88 信号转导通路激活 NF-κB,进一步增强 JEG-3 细胞的增殖、迁移和侵袭。