Department of Obstetrical, The First People's Hospital of Jining, Jining, China.
Eur Rev Med Pharmacol Sci. 2018 Oct;22(20):6583-6590. doi: 10.26355/eurrev_201810_16132.
The role of miR-182-5p in preeclampsia was studied, and its mechanism was also explored.
Fifty patients with preeclampsia were assigned to the study group and 50 normal pregnant women to the control group. The age, weight, blood pressure, urinary protein, and weight of newborns were compared between the two groups. The placental tissues of the above-mentioned subjects were collected, and quantitative Reverse Transcriptase-Polymerase Chain Reaction (qRT-PCR) assay was used to detect the expression of miR-182-5p. MiR-182-5p was overexpressed or knocked down using a cell transfection assay in HTR-8/SVneov cell, which is a kind of human chorionic trophoblast cell. Changes in cell migration and invasiveness before and after transfection were determined by wound healing test and transwell assay, respectively. Western blot was performed to analyze the change of RND3 protein level before and after transfection. The biological prediction of the relationship between miR-182-5p and RND3 was performed and a dual luciferase reporter gene experiment was designed to verify the results. Finally, a rescue experiment was conducted to investigate whether RND3 could affect the role of miRNA-182-5p in the capacity of cell migration and invasion.
Preeclampsia patients had higher systolic blood pressure, diastolic blood pressure, and urinary protein than normal pregnant women, while neonatal weight decreased compared with normal pregnant women. MiRNA-182-5p was highly expressed in placental tissues of patients with preeclampsia. After miRNA-182-5p was overexpressed, the migration and invasion of HTR-8/SVneo cells were significantly attenuated, and the mRNA and protein levels of RND3 were markedly downregulated, and vice versa. The dual luciferase reporting assay confirmed that miRNA-182-5p could bind to 3'UTR of RND3. In addition, the results of rescue experiment showed that overexpressing miRNA-182-5p could markedly inhibit the migration and invasion of HTR-8/SVneo cells; however, when RND3 was simultaneously overexpressed, the inhibitory effect of miRNA-182-5p was partially reversed.
The highly expressed miRNA-182-5p in patients with preeclampsia promoted the development of preeclampsia, the possible mechanism of which might be that the increased miRNA-182-5p expression could inhibit the migratory and invasive ability of trophoblast cells through targeted degrading RND3 protein.
研究 miR-182-5p 在子痫前期中的作用，并探讨其机制。
将 50 例子痫前期患者纳入研究组，50 例正常妊娠孕妇纳入对照组。比较两组患者的年龄、体重、血压、尿蛋白、新生儿体重。收集上述患者的胎盘组织，采用实时定量逆转录聚合酶链反应（qRT-PCR）检测 miR-182-5p 的表达。采用细胞转染实验在 HTR-8/SVneov 细胞（一种人绒毛滋养层细胞）中转染 miR-182-5p，观察细胞迁移和侵袭能力的变化。通过划痕实验和 Transwell 实验分别检测转染前后细胞迁移和侵袭能力的变化。采用 Western blot 检测转染前后 RND3 蛋白水平的变化。对 miR-182-5p 与 RND3 之间的关系进行生物预测，并设计双荧光素酶报告基因实验验证结果。最后，进行挽救实验，以探讨 RND3 是否能影响 miRNA-182-5p 对细胞迁移和侵袭能力的作用。
子痫前期患者的收缩压、舒张压和尿蛋白均高于正常孕妇，而新生儿体重低于正常孕妇。子痫前期患者胎盘组织中 miR-182-5p 表达水平升高。miR-182-5p 过表达后，HTR-8/SVneo 细胞的迁移和侵袭能力明显减弱，RND3 的 mRNA 和蛋白水平明显下调，反之亦然。双荧光素酶报告基因实验证实 miR-182-5p 可与 RND3 的 3'UTR 结合。此外，挽救实验结果表明，过表达 miR-182-5p 可显著抑制 HTR-8/SVneo 细胞的迁移和侵袭；然而，当同时过表达 RND3 时，miR-182-5p 的抑制作用部分逆转。
子痫前期患者中高表达的 miR-182-5p 促进了子痫前期的发生发展，其可能的机制是，miR-182-5p 表达增加通过靶向降解 RND3 蛋白抑制滋养层细胞的迁移和侵袭能力。
