Department of Obstetrics, Affiliated Hangzhou First People's Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang 310006, P.R. China.
Mol Med Rep. 2021 May;23(5). doi: 10.3892/mmr.2021.11958. Epub 2021 Mar 24.
Pregnancy‑induced hypertension is often accompanied by preeclampsia. The present study investigated whether microRNA (miR)‑27b‑3p affected the occurrence of preeclampsia by regulating the function of endothelial cells. Expressions levels of miR‑27b‑3p and ATPase plasma membrane Ca transporting 1 (ATP2B1) were determined using reverse‑transcription quantitative PCR. miR‑27b‑3p targeting ATP2B1 was predicted using bioinformatics and further confirmed by dual‑luciferase reporter assays. Cell Counting Kit‑8, Transwell and Matrigel tube formation assays were performed to detect the effects of miR‑27b‑3p on proliferation, migration and tube formation of human umbilical vein endothelial cells (HUVECs), respectively. Moreover, HTR8/SVneos cells were co‑cultured with HUVECs to detect the invasion of trophoblast cells, and the expression levels of vascular endothelial growth factor (VEGF), matrix metalloproteinase (MMP)‑2 and MMP‑9 of HUVECs and HTR8/SVneos were detected by western blotting. Expression levels of miR‑27b‑3p were upregulated in the serum of patients with hypertension and preeclampsia, which could target and regulate the expression of ATP2B1. The expression levels of miR‑27b‑3p were increased and those of ATP2B1 were reduced in HUVECs from hypertensive serums. Moreover, miR‑27b‑3p mimics reduced the expression level of ATP2B1, and miR‑27b‑3p inhibitor reversed the effect of hypertensive serum on ATP2B1 expression. Furthermore, patients with hypertension showed increased endothelial dysfunction, reduced trophoblastic invasion and the expressions of VEGF, MMP‑2 and MMP‑9, and miR‑27b‑3p mimics and silencing of ATP2B1 produced similar results to HUVECs. The miR‑27b‑3p inhibitor reversed the effect of hypertensive serum, and silencing of ATP2B1 inhibited the improvement of miR‑27b‑3p inhibitor to HUVECs and HTR‑8/SVneo cells in proliferation, migration and tube formation. The current findings suggested that miR‑27b‑3p promoted proliferation, migration and tube formation of HUVECs and enhanced invasion of trophoblast cells, via regulation of ATP2B1. Thus, miR‑27b‑3p could be considered as a molecular risk factor in the pathogenesis and development of preeclampsia.
妊娠高血压常伴有子痫前期。本研究探讨了 microRNA(miR)-27b-3p 是否通过调节内皮细胞的功能来影响子痫前期的发生。采用逆转录定量 PCR 测定 miR-27b-3p 和 ATP 酶质膜 Ca 转运 1(ATP2B1)的表达水平。使用生物信息学预测 miR-27b-3p 靶向 ATP2B1,并通过双荧光素酶报告基因检测进一步证实。通过细胞计数试剂盒-8 检测、Transwell 检测和 Matrigel 管形成检测分别检测 miR-27b-3p 对人脐静脉内皮细胞(HUVEC)增殖、迁移和管形成的影响。此外,将 HTR8/SVneos 细胞与 HUVEC 共培养,检测滋养层细胞的侵袭,Western blot 检测 HUVEC 和 HTR8/SVneos 的血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)-2 和 MMP-9 的表达水平。高血压和子痫前期患者血清中 miR-27b-3p 的表达上调,可靶向调节 ATP2B1 的表达。高血压血清中 HUVEC 中 miR-27b-3p 的表达上调,ATP2B1 的表达下调。此外,miR-27b-3p 模拟物降低了 ATP2B1 的表达水平,而高血压血清对 ATP2B1 表达的抑制作用被 miR-27b-3p 抑制剂逆转。此外,高血压患者表现出内皮功能障碍增加、滋养层细胞侵袭减少以及 VEGF、MMP-2 和 MMP-9 的表达减少,miR-27b-3p 模拟物和沉默 ATP2B1 产生了与 HUVEC 类似的结果。miR-27b-3p 抑制剂逆转了高血压血清的作用,而沉默 ATP2B1 抑制了 miR-27b-3p 抑制剂对 HUVEC 和 HTR-8/SVneo 细胞增殖、迁移和管形成的改善作用。目前的研究结果表明,miR-27b-3p 通过调节 ATP2B1 促进 HUVEC 的增殖、迁移和管形成,并增强滋养层细胞的侵袭。因此,miR-27b-3p 可作为子痫前期发病和发展的分子危险因素。