Maternity Department, Ningbo Women & Children's Hospital, Ningbo City, Zhejiang Province, China.
J Clin Lab Anal. 2022 Jul;36(7):e24549. doi: 10.1002/jcla.24549. Epub 2022 Jun 13.
Mast cells regulate the process of preeclampsia (PE). Since we previously identified mast cells specifically expressing miR-181a-5p in the placenta of PE patients, it is plausible to examine the effect and mechanism of mast cell-derived exosomal miR-181a-5p on trophoblast cells.
The miR-181a-5p and YY1 levels were determined by quantitative real-time reverse transcription-polymerase chain reaction. Exosomes were identified by transmission electron microscopy, Western blot, and PKH-26 labeling. Mast cells or trophoblast cell malignant phenotype were detected using 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide, wound healing, and Transwell assays. Quantification of YY1 and metastasis-related proteins was performed using Western blot. TargetScan, JASPAR, dual-luciferase reporter genes, and chromatin immunoprecipitation were exploited to verify the relationship between miR-181a-5p, YY1, and MMP-9.
MiR-181a-5p was overexpressed in mast cells of PE patients. Overexpressed miR-181a-5p restrained mast cell viability. Mast cell exosomes were successfully isolated, containing high expressions of CD63 and HSP70 and low expression of Calnexin and could be transported to the cytoplasm of trophoblast cells. Mast cell exosomes attenuated the viability, migration, and invasion of HTR-8/SVneo cells, inhibited YY1, N-cadherin, Vimentin, and MMP-9 protein expressions, and promoted E-cadherin protein expression. The effect of exosomes was enhanced by miR-181a-5p mimic but was reversed by miR-181a-5p inhibitor. MiR-181a-5p targeted YY1 which bound to the MMP-9 promoter. Overexpressed YY1 in HTR-8/SVneo cells accelerated the malignant phenotype of the cells and reversed the regulatory effects of exosomal miR-181a-5p.
Mast cell-derived exosomal miR-181a-5p modulates HTR-8/SVneo cell viability, migration, and invasion via YY1/MMP-9.
肥大细胞调节子痫前期（PE）的发生过程。由于我们之前在 PE 患者的胎盘组织中鉴定出了特异性表达 miR-181a-5p 的肥大细胞，因此研究肥大细胞来源的外泌体 miR-181a-5p 对滋养层细胞的作用和机制是合理的。
通过实时定量逆转录聚合酶链反应测定 miR-181a-5p 和 YY1 的水平。通过透射电子显微镜、Western blot 和 PKH-26 标记鉴定外泌体。通过 3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴盐、划痕愈合和 Transwell 测定法检测肥大细胞或滋养层细胞的恶性表型。通过 Western blot 定量测定 YY1 和转移相关蛋白。利用 TargetScan、JASPAR、双荧光素酶报告基因和染色质免疫沉淀实验验证 miR-181a-5p、YY1 和 MMP-9 之间的关系。
PE 患者的肥大细胞中 miR-181a-5p 过表达。过表达的 miR-181a-5p 抑制了肥大细胞的活力。成功分离了肥大细胞外泌体，其中高表达 CD63 和 HSP70，低表达 Calnexin，并能被转运到滋养层细胞的细胞质中。肥大细胞外泌体减弱了 HTR-8/SVneo 细胞的活力、迁移和侵袭能力，抑制了 YY1、N-钙黏蛋白、波形蛋白和 MMP-9 蛋白的表达，促进了 E-钙黏蛋白的表达。外泌体的作用通过 miR-181a-5p 模拟物增强，但通过 miR-181a-5p 抑制剂逆转。miR-181a-5p 靶向与 MMP-9 启动子结合的 YY1。在 HTR-8/SVneo 细胞中转染过表达的 YY1 加速了细胞的恶性表型，并逆转了外泌体 miR-181a-5p 的调节作用。
肥大细胞来源的外泌体 miR-181a-5p 通过 YY1/MMP-9 调节 HTR-8/SVneo 细胞的活力、迁移和侵袭。
