Center for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, Shandong,China; Center for Reproductive Medicine, Jinan Central Hospital , Cheeloo College of Medicine ,Shandong University, Jinan, Shandong,China.
Center for Reproductive Medicine, Cheeloo College of Medicine, Shandong University, Jinan, Shandong,China; Department of Obstetrics and Gynecology, Qingdao Municipal Hospital, Shandong University, Qingdao, China.
J Reprod Immunol. 2020 Aug;140:103133. doi: 10.1016/j.jri.2020.103133. Epub 2020 Apr 18.
Recurrent spontaneous abortion (RSA) can be attributed to multiple factors, and abnormal invasion and apoptosis of trophoblast cells have attracted extensive attention. Our study aimed to investigate the expression of PTEN and miRNAs with potential regulatory relationships in the placental villi of RSA patients. Nineteen RSA patients and sixteen healthy women at reproductive age undergoing induced abortion (IA) were enrolled in the present study. The expression of PTEN and miRNAs were investigated using real-time polymerase chain reaction (PCR) and Western blotting, further verification between PTEN and potential miRNAs used cell culture and transfection, and luciferase activity assays were used to determine whether PTEN is directly regulated by potential miRNA. The results indicated that both PTEN mRNA and protein expression levels were upregulated in RSA patients, but a significant difference was only observed in protein expression level (p < 0.001). Through real-time PCR pre-scanning, the results of nine potential miRNAs revealed three significantly upregulated miRNAs (miR-494, miR-146a, and miR-21) and one significantly downregulated miRNA (miR-19b). The results of further verification regarding miR-19b and miR-494 suggested that upregulated miR-19b, cooperating with downregulated miRNA-494, could inhibit PTEN expression. In conclusion, the findings suggest that the overexpression of PTEN plays an important role in the pathogenesis of RSA, with miR-19b directly regulating PTEN and working with miR-494, all of which participating in abnormal effects of villous' trophoblastic cell may be a critical event.
复发性自然流产（RSA）可归因于多种因素，滋养细胞的异常侵袭和凋亡引起了广泛关注。本研究旨在探讨 RSA 患者胎盘绒毛中具有潜在调节关系的 PTEN 和 miRNA 的表达。本研究纳入了 19 例 RSA 患者和 16 例年龄匹配的因非意愿妊娠行人工流产（IA）的健康对照者。采用实时聚合酶链反应（PCR）和 Western blot 检测 PTEN 和 miRNA 的表达，通过细胞培养和转染进一步验证 PTEN 与潜在 miRNA 之间的关系，采用荧光素酶活性测定确定 PTEN 是否受潜在 miRNA 的直接调控。结果表明，RSA 患者中 PTEN mRNA 和蛋白表达水平均上调，但仅在蛋白表达水平上差异有统计学意义（p < 0.001）。通过实时 PCR 预扫描，9 种潜在 miRNA 的结果显示有 3 种 miRNA（miR-494、miR-146a 和 miR-21）显著上调，1 种 miRNA（miR-19b）显著下调。进一步验证 miR-19b 和 miR-494 的结果表明，上调的 miR-19b 与下调的 miR-494 协同作用可抑制 PTEN 表达。总之，研究结果提示 PTEN 的过度表达在 RSA 的发病机制中起重要作用，miR-19b 直接调控 PTEN 并与 miR-494 协同作用，共同参与绒毛滋养细胞的异常效应可能是一个关键事件。
