Obstetrics Department, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, P.R. China.
Gynecologic Oncology Department, Beijing Obstetrics and Gynecology Hospital, Capital Medical University, Beijing 100026, P.R. China.
Biosci Rep. 2019 Dec 20;39(12). doi: 10.1042/BSR20191271.
The present study aimed to investigate the underlying mechanism of miR-126a-3p in the proliferation, migration and invasion of trophoblast cells in pre-eclampsia-like rats by targeting A Disintegrin and Metalloprotease 9 (ADAM9). First, the interaction between miR-126a-3p and ADAM9 was confirmed via biochemical assays. Placental tissues and trophoblast cells were then obtained. RNA in situ hybridization was performed in order to detect miR-126a-3p expression in the placenta. Subsequently, a series of biological assays, including reverse transcription-quantitative PCR (RT-qPCR), Western blotting, MTT assay, apoptosis assay, cell cycle assay, wound healing assay and transwell assay were adopted in order to determine the cell proliferation, cell cycle distribution, apoptotic rate, and migration and invasion of trophoblast cells in each group. The results revealed that miR-126a-3p was down-regulated in the placenta of pre-eclampsia-like rats. In vivo experiments' results indicated that miR-126a-3p could inhibit ADAM9 expression, and induce cyclin D1, Matrix metalloproteinase (MMP) 2 (MMP-2), MMP-9 expression. MTT, apoptosis and cell cycle assay results revealed that trophoblast cells transfected with miR-126a-3p mimic or si-ADAM9 exhibited higher proliferative activity and a lower apoptotic rate compared with the blank group (all P<0.05). The wound healing assay and transwell assay results confirmed that, compared with the blank group, the migration and invasion ability of trophoblast cells in the miR-126a-3p mimic group and small interfering RNA (siRNA)-ADAM9 group were significantly increased (all P<0.05). Conversely, miR-126a-3p inhibitor treatment revealed the opposite effect (all P<0.05). In conclusion, the present study demonstrated that miR-126a-3p could enhance proliferation, migration and invasion, but decrease the apoptosis rate of trophoblast cells in pre-eclampsia-like rats through targeting ADAM9.
本研究旨在通过靶向解整合素金属蛋白酶 9（ADAM9）探讨 miR-126a-3p 在子痫前期样大鼠滋养细胞增殖、迁移和侵袭中的作用机制。首先，通过生化实验验证了 miR-126a-3p 与 ADAM9 的相互作用。然后获取胎盘组织和滋养细胞，采用 RNA 原位杂交检测 miR-126a-3p 在胎盘组织中的表达。随后，采用逆转录-定量聚合酶链反应（RT-qPCR）、Western blot、MTT 实验、细胞凋亡实验、细胞周期实验、划痕愈合实验和 Transwell 实验检测各组滋养细胞的增殖、细胞周期分布、凋亡率、迁移和侵袭能力。结果表明，子痫前期样大鼠胎盘中 miR-126a-3p 表达下调。体内实验结果表明，miR-126a-3p 可抑制 ADAM9 表达，并诱导细胞周期蛋白 D1、基质金属蛋白酶 2（MMP-2）、MMP-9 表达。MTT、凋亡和细胞周期实验结果显示，转染 miR-126a-3p 模拟物或 si-ADAM9 的滋养细胞表现出更高的增殖活性和更低的凋亡率，与空白组相比差异均有统计学意义（均 P<0.05）。划痕愈合实验和 Transwell 实验结果证实，与空白组相比，miR-126a-3p 模拟物组和 si-ADAM9 组滋养细胞的迁移和侵袭能力显著增强（均 P<0.05）。相反，miR-126a-3p 抑制剂处理则呈现相反的效果（均 P<0.05）。综上所述，本研究表明 miR-126a-3p 通过靶向 ADAM9 增强子痫前期样大鼠滋养细胞的增殖、迁移和侵袭能力，同时降低其凋亡率。
