Department of Obstetrics and Gynecology, Department of Fetal Medicine and Prenatal Diagnosis, Key Laboratory for Major Obstetric Diseases of Guangdong Province, The Third Affiliated Hospital of Guangzhou Medical University, 63 Duobao Road, Liwan District, Guangzhou, China.
The University of Hong Kong Shenzhen Key Laboratory of Fertility Regulation, The University of Hong Kong-Shenzhen Hospital, Shenzhen, China.
Reprod Sci. 2022 Apr;29(4):1296-1304. doi: 10.1007/s43032-021-00663-7. Epub 2021 Aug 30.
Extravillous cytotrophoblast (EVCT) is responsible for trophoblast invasion, which is important during placentation. Dysregulation of the process leads to pregnancy complications. S-nitrosylation of proteins is associated with cell invasion in many cell types. Adrenomedullin (ADM), a polypeptide expressed abundantly in the first-trimester placentas, induces EVCT invasion by upregulation of protein S-nitrosylation. This study aimed to identify the S-nitrosylated proteins induced by ADM in the JEG-3 placental cells. By using affinity chromatography followed by mass spectrometric analysis, tubulin, enolase, eukaryotic translation initiation factor 4A1, actin, annexin II (ANX II), and glyceraldehyde 3-phosphate dehydrogenaseprotein-1 were found to be S-nitrosylated by ADM. In vitro treatment with ADM or S-Nitrosoglutathione (GSNO) significantly increased the ANX II surface expression, but not its total expression in the JEG-3 cells. Translocation of ANX II to cell surface has been reported to act as a cell surface receptor to plasmin, plasminogen, and tissue plasminogen activator (tPA), thereby stimulating cell invasion and migration. However, in this study, ADM-induced surface expression of ANX II in the JEG-3 cells was not associated with changes in the secretory and membrane-bound tPA activities. Future studies are required to understand the roles of surface expression of S-nitrosylated ANX II on trophoblast functions. To conclude, this study provided evidences that ADM regulated the nitric oxide signaling pathway and modulated trophoblast invasion.
绒毛外滋养细胞（EVCT）负责滋养细胞浸润，这在胎盘形成过程中很重要。该过程的失调会导致妊娠并发症。许多细胞类型中的蛋白质的 S-亚硝基化与细胞浸润有关。在孕早期胎盘大量表达的多肽类物质肾上腺髓质素（ADM）通过上调蛋白质 S-亚硝基化诱导 EVCT 浸润。本研究旨在鉴定 ADM 在 JEG-3 胎盘细胞中诱导的 S-亚硝基化蛋白。通过使用亲和层析，然后进行质谱分析，发现微管蛋白、烯醇酶、真核翻译起始因子 4A1、肌动蛋白、膜联蛋白 II（ANX II）和甘油醛 3-磷酸脱氢酶蛋白-1 被 ADM 进行 S-亚硝基化。在体外用 ADM 或 S-亚硝基谷胱甘肽（GSNO）处理可显著增加 JEG-3 细胞中 ANX II 的表面表达，但不增加其总表达。据报道，膜联蛋白 II 向细胞表面的易位作为纤维蛋白溶酶、纤溶酶原和组织型纤溶酶原激活剂（tPA）的细胞表面受体，从而刺激细胞浸润和迁移。然而，在本研究中，ADM 诱导的 JEG-3 细胞中 ANX II 的表面表达与分泌型和膜结合型 tPA 活性的变化无关。需要进一步的研究来了解 S-亚硝基化的 ANX II 表面表达在滋养细胞功能中的作用。总之，本研究提供了证据表明 ADM 调节了一氧化氮信号通路并调节了滋养细胞浸润。
