Department of Obstetrics, Shandong Provincial Hospital Affiliated to Shandong First Medical University, 324 Jingwu Road, Jinan, 250021, China.
Key Laboratory of Birth Regulation and Control Technology of National Health and Family Planning Commission of China, Maternal Child Health Hospital of Shandong Province, 328 Jingshi East Road, Jinan, China.
BMC Pregnancy Childbirth. 2021 Dec 20;21(1):837. doi: 10.1186/s12884-021-04069-w.
Preeclampsia (PE) is a major cause of maternal and perinatal morbidity and mortality. Studies on the role of microRNAs (miRNAs), in the pathogenesis of PE through their effects on trophoblast function have been reported, but roles for some miRNAs including miR-513c-5p, have not been identified. We aimed to evaluate potential miRNA candidates that regulate the LRP6 mRNAand to elucidate the possible mechanism in PE. Potential miRNAs were selected by bioinformatics analysis, PCR of placenta tissues and dual luciferase reporter assay of HTR-8/SVneo cells.
A bioinformatics analysis (Gene Expression Omnibus, GEO; miRWalk) was performed to screen the possible miRNAs that participate in the pathology of PE. Placentas from patients with PE and women with a normal pregnancy were collected to detect the expression of predicted miRNAs by RT-qPCR. A dual luciferase reporter assay was used to test the binding of the potential miRNAs to LRP6. The effects of miR-513c-5p on the biological functions of HTR-8/SVneo cells were further evaluated by performing EdU staining, flow cytometry, wound healing assays and Transwell assays.
GEO and miRWalk predicted 16 miRNAs that might target LRP6. Hsa-miR-371a-5p, hsa-miR-513c-5p, hsa-miR-126-3p, hsa-miR-145-5p, hsa-miR-193b-5p and hsa-miR-296-5p were 6 miRNAs upregulated in the PE placenta. LRP6 was downregulated in patients with PE compared to normal women. miR-513c-5p mimics inhibited LRP6 expression in HTR-8/SVneo cells, and LRP6 is the target gene of miR-513c-5p. miR-513c-5p mimics also inhibited invasion, migration and proliferation of HTR-8/SVneo cells but promoted their apoptosis.
Our study reveals that overexpression of placenta miR-513c-5p is involved in PE by regulating the biological functions of trophoblasts through the inhibition of LRP6.
子痫前期（PE）是孕产妇和围生儿发病率和死亡率的主要原因。已有研究表明，microRNAs（miRNAs）通过影响滋养层功能在 PE 的发病机制中起作用，但包括 miR-513c-5p 在内的一些 miRNAs 的作用尚未确定。我们旨在评估调节 LRP6 mRNA 的潜在 miRNA 候选物，并阐明 PE 中的可能机制。通过生物信息学分析、胎盘组织 PCR 和 HTR-8/SVneo 细胞双荧光素酶报告基因检测来选择潜在的 miRNA。
通过生物信息学分析（Gene Expression Omnibus，GEO；miRWalk）筛选可能参与 PE 病理的 miRNA。收集 PE 患者和正常妊娠妇女的胎盘，通过 RT-qPCR 检测预测 miRNA 的表达。使用双荧光素酶报告基因检测潜在 miRNA 与 LRP6 的结合。通过 EdU 染色、流式细胞术、划痕愈合实验和 Transwell 实验进一步评估 miR-513c-5p 对 HTR-8/SVneo 细胞生物学功能的影响。
GEO 和 miRWalk 预测了 16 个可能靶向 LRP6 的 miRNA。hsa-miR-371a-5p、hsa-miR-513c-5p、hsa-miR-126-3p、hsa-miR-145-5p、hsa-miR-193b-5p 和 hsa-miR-296-5p 是 6 个在 PE 胎盘中上调的 miRNA。与正常女性相比，PE 患者的 LRP6 表达下调。miR-513c-5p 模拟物抑制 HTR-8/SVneo 细胞中的 LRP6 表达，LRP6 是 miR-513c-5p 的靶基因。miR-513c-5p 模拟物还抑制 HTR-8/SVneo 细胞的侵袭、迁移和增殖，但促进其凋亡。
本研究表明，胎盘 miR-513c-5p 的过度表达通过抑制 LRP6 调节滋养层细胞的生物学功能参与 PE 的发生。
