Highly expressed miR-182-5p can promote preeclampsia progression by degrading RND3 and inhibiting HTR-8/SVneo cell invasion
Highly expressed miR-182-5p can promote preeclampsia progression by degrading RND3 and inhibiting HTR-8/SVneo cell invasion
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DOI：
10.26355/eurrev_201810_16132
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发表时间：
2018-10-01
影响因子：
3.3
通讯作者：
Wu, J-L
中科院分区：
文献类型：
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作者：
Fang, Y-N;Huang, Z-L;Wu, J-L
目的：研究miR-182-5p在子痫前期中的作用，并探讨其机制。 患者与方法：将50例子痫前期患者纳入研究组，将50例正常孕妇纳入对照组。比较两组新生儿的年龄、体重、血压、尿蛋白、体重。收集上述受试者的胎盘组织，采用定量逆转录酶-聚合酶链反应（qRT-PCR）法检测miR-182-5p的表达量。使用细胞转染测定在 HTR-8/SVneov 细胞（一种人绒毛膜滋养层细胞）中过度表达或敲除 MiR-182-5p。分别通过伤口愈合试验和Transwell实验测定转染前后细胞迁移和侵袭力的变化。 Western blot分析转染前后RND3蛋白水平的变化。对miR-182-5p与RND3之间的关系进行生物学预测，并设计双荧光素酶报告基因实验来验证结果。最后进行挽救实验，探讨RND3是否会影响miRNA-182-5p在细胞迁移和侵袭能力中的作用。结果：子痫前期患者的收缩压、舒张压和尿蛋白均高于正常孕妇，而新生儿体重较正常孕妇有所下降。 miRNA-182-5p在子痫前期患者的胎盘组织中高表达。 miRNA-182-5p过表达后，HTR-8/SVneo细胞的迁移和侵袭能力显着减弱，RND3的mRNA和蛋白水平显着下调，反之亦然。双荧光素酶报告测定证实 miRNA-182-5p 可以与 RND3 的 3'UTR 结合。此外，拯救实验结果表明，过表达miRNA-182-5p能够显着抑制HTR-8/SVneo细胞的迁移和侵袭；结论：子痫前期患者高表达的miRNA-182-5p促进了子痫前期的发生，其可能机制是miRNA-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。
OBJECTIVE: The role of miR-182-5p in preeclampsia was studied, and its mechanism was also explored.PATIENTS AND METHODS: 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.RESULTS: 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.CONCLUSIONS: 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.