Department of Obstetrics and Gynecology, The Eighth Hospital of Wuhan, Wuhan, China.
Department of Cardiology, WuHan FangTai Hospital, Wuhan, China.
Bioengineered. 2022 May;13(5):12338-12349. doi: 10.1080/21655979.2022.2073655.
The level of miR‑135b-5p is lower in patients with preeclampsia (PE) superimposed on chronic hypertension than in healthy controls. However, the function of miR‑135b-5p in PE progression remains unknown. In the present study, we investigated the role of miR‑135b-5p in PE development and its possible mechanism for the first time. HTR8/SVneo cells (trophoblast cell line) were exposed to hypoxia/reoxygenation (H/R) to mimic PE . Hypoxia-inducible factor-1α (HIF-1α), forkhead box O3A (FOXO3a), and miR-135b-5p levels were measured using Real-time PCR. Cell proliferation, apoptosis and migration/invasion were evaluated using the Cell Counting Kit-8 (CCK-8), flow cytometry and transwell assays, respectively. Real-time PCR and Western blotting were performed to determine the levels of several pro- and anti-angiogenic factors. The binding of miR-135b-5p to the PIK3R2-3' untranslated region (3'UTR) was confirmed by bioinformatics analysis and a dual-luciferase reporter assay. H/R exposure greatly upregulated HIF-1α, FOXO3a, and PIK3R2 levels, while downregulating miR-135b-5p levels in HTR8/SVneo cells. H/R exposure resulted in the inhibition of proliferation, migration, invasion, angiogenesis, and the induction of apoptosis. MiR-135b-5p overexpression reversed the effects of H/R on trophoblast cell function, while miR-135b-5p knockdown enhanced the effects. PIK3R2 knockdown had similar effects as miR-135b-5p overexpression on proliferation, apoptosis and angiogenesis. The effect of miR-135b-5p overexpression on H/R-exposed cells was enhanced by PIK3R2 knockdown. MiR-135b-5p downregulated PIK3R2 expression by pairing with its 3'UTR. Therefore, miR-135b-5p may regulate trophoblast function by targeting PIK3R2 in PE and could serve as a novel therapeutic target for PE.
子痫前期（PE）合并慢性高血压患者的 miR-135b-5p 水平低于健康对照者。然而，miR-135b-5p 在 PE 进展中的作用尚不清楚。本研究首次探讨了 miR-135b-5p 在 PE 发病机制中的作用及其可能的机制。用缺氧/复氧（H/R）处理 HTR8/SVneo 细胞（滋养层细胞系）模拟 PE。采用实时 PCR 检测缺氧诱导因子 1α（HIF-1α）、叉头框 O3A（FOXO3a）和 miR-135b-5p 的水平。采用细胞计数试剂盒-8（CCK-8）、流式细胞术和 Transwell 测定法分别评估细胞增殖、凋亡和迁移/侵袭。采用实时 PCR 和 Western blot 测定法测定几种促血管生成和抗血管生成因子的水平。通过生物信息学分析和双荧光素酶报告基因测定证实 miR-135b-5p 与 PIK3R2-3'UTR 的结合。H/R 暴露可显著上调 HTR8/SVneo 细胞中 HIF-1α、FOXO3a 和 PIK3R2 的水平，同时下调 miR-135b-5p 的水平。H/R 暴露可抑制增殖、迁移、侵袭、血管生成，并诱导凋亡。miR-135b-5p 过表达可逆转 H/R 对滋养层细胞功能的影响，而 miR-135b-5p 敲低则增强了这种作用。PIK3R2 敲低对增殖、凋亡和血管生成的影响与 miR-135b-5p 过表达相似。PIK3R2 敲低增强了 miR-135b-5p 过表达对 H/R 暴露细胞的作用。miR-135b-5p 通过与其 3'UTR 配对下调 PIK3R2 的表达。因此，miR-135b-5p 可能通过靶向 PIK3R2 调节 PE 中的滋养层功能，可作为 PE 的一种新的治疗靶点。
