Clinical Center of Reproductive Medicine, Lianyungang Maternal and Child Health Hospital, Kangda College of Nanjing Medical University, Lianyungang, China.
State Key Laboratory of Reproductive Medicine, Clinical Center of Reproductive Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Mol Genet Genomic Med. 2023 Sep;11(9):e2220. doi: 10.1002/mgg3.2220. Epub 2023 Jun 8.
Enolase 1 (ENO1) is a metabolic enzyme which participates in pyruvate synthesis and ATP production in cells. Previously, differential expression of ENO1 was discovered in villous tissues between recurrent miscarriage and induced abortion. This study was designed to explore whether ENO1 influences the proliferation and invasion of villous trophoblasts and the related molecular mechanisms.
First, ENO1 expression in placental villus tissues collected from recurrent miscarriage (RM) patients and women for induced abortion as well as in trophoblast-derived cell lines was detected by RT-qPCR and western blotting. ENO1 localization and expression in villus tissues were further confirmed through immunohistochemistry staining. Then, the effects of ENO1 downregulation on trophoblast Bewo cell proliferation, migration, invasion, and epithelial-mesenchymal transition (EMT) process were evaluated by CCK-8 assay, transwell assay, and western blotting. As for the regulatory mechanism of ENO1, the expression of COX-2, c-Myc and cyclin D1 in Bewo cells after ENO1 knockdown was finally evaluated by RT-qPCR and western blotting.
ENO1 was mainly localized in the cytoplasm, with very small amounts in the nucleus of trophoblast cells. ENO1 expression in the villi tissues of RM patients was significantly increased, when compared with the villous tissues of healthy controls. Furthermore, Bewo cells, a trophoblast cell line with relatively higher expression of ENO1, was used to downregulate the ENO1 expression by ENO1-siRNA transfection. ENO1 knockdown significantly facilitated Bewo cell growth, EMT process, migration, and invasion. ENO1 silencing markedly elevated COX-2, c-Myc, and cyclin D1 expression.
ENO1 may participate in the development of RM via suppressing the growth and invasion of villous trophoblasts via reducing the expression of COX-2, c-Myc, and cyclin D1.
烯醇化酶 1（ENO1）是一种代谢酶，参与细胞内丙酮酸的合成和 ATP 的产生。先前，在复发性流产和人工流产的绒毛组织中发现 ENO1 的差异表达。本研究旨在探讨 ENO1 是否影响绒毛滋养层的增殖和侵袭及其相关的分子机制。
首先，通过 RT-qPCR 和 Western blot 检测复发性流产（RM）患者和人工流产妇女胎盘绒毛组织以及滋养层来源细胞系中 ENO1 的表达。通过免疫组织化学染色进一步证实 ENO1 在绒毛组织中的定位和表达。然后，通过 CCK-8 测定、Transwell 测定和 Western blot 评估 ENO1 下调对 Bewo 细胞增殖、迁移、侵袭和上皮-间充质转化（EMT）过程的影响。至于 ENO1 的调节机制，最后通过 RT-qPCR 和 Western blot 评估 ENO1 敲低后 Bewo 细胞中 COX-2、c-Myc 和 cyclin D1 的表达。
ENO1 主要定位于细胞质中，细胞核中含量很少。与健康对照组相比，RM 患者的绒毛组织中 ENO1 的表达明显增加。此外，Bewo 细胞是一种 ENO1 表达相对较高的滋养层细胞系，通过 ENO1-siRNA 转染下调 ENO1 表达。ENO1 敲低显著促进 Bewo 细胞生长、EMT 过程、迁移和侵袭。ENO1 沉默显著提高 COX-2、c-Myc 和 cyclin D1 的表达。
ENO1 可能通过降低 COX-2、c-Myc 和 cyclin D1 的表达，抑制绒毛滋养层的生长和侵袭，参与 RM 的发生。
