MicroRNA-135b-5p regulates trophoblast cell function by targeting phosphoinositide-3-kinase regulatory subunit 2 in preeclampsia

Abstract

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 in vitro . 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 (3ʹUTR) 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 3ʹUTR. Therefore, miR-135b-5p may regulate trophoblast function by targeting PIK3R2 in PE and could serve as a novel therapeutic target for PE.

Extracellular vesicles derived from Akkermansia muciniphila promote placentation and mitigate preeclampsia in a mouse model

“…The translocation of AmEVs in the placenta and the remission of PE symptoms in AmEVs‐pretreated mice prompted us to investigate the potential mechanism of AmEVs in human cell line. Accordingly, we employed the human first‐trimester trophoblast HTR‐8/SVneo cell line to establish a hypoxia/reoxygenation (H/R) model to mimic the state of PE trophoblast based on previous studies (Zhang & Zhang, 2022 ). The cell proliferation rate was significantly higher in groups pretreated with 10 or 20 μg/mL AmEVs than in the H/R group (Figure 7a ).…”

“…The cells were grown in a humidified incubator at 37°C with 5% CO 2 . To establish a trophoblast cell model of PE, the HTR‐8/SVneo cells were cultured at 37°C for 8 h under hypoxic conditions (2% O 2 ), followed by 16 h of reoxygenation (H/R) under 5% CO 2 /air; the cells were grown under 5% CO 2 /air at 37°C and served as controls (Zhang & Zhang, 2022 ). To test the impact of AmEVs, cells were pretreated with different concentrations (1, 10, 20 or 50 μg/mL) of AmEVs in a fresh culture medium for 24 h, and then subjected to 8 h of hypoxia and 16 h of reoxygenation.…”

6-Gingerol alleviates placental injury in preeclampsia by inhibiting oxidative stress via BNIP3/LC3 signaling-mediated trophoblast mitophagy

“…Therefore, as a NO synthase inhibitor, L-NAME gives rise to oxidative stress injury in the placenta, which has been demonstrated in the present study. In addition, many studies have been carried out on trophoblast cell models to elucidate the origin of oxidative stress, specifically H/R-treated HTR8/SVneo cells (Gu et al, 2022;Zhang and Zhang, 2022). Several studies suggested that placental lipid peroxides increased and placental antioxidant protective effects decreased in PE (Vaka et al, 2021).…”

LncRNA GAS5 Modulates the Progression of Glioma Through Repressing miR-135b-5p and Upregulating APC

“…The miRNA and mRNA expression levels were normalized against U6 and GAPDH, respectively. 31,[41][42][43][44] Reaction conditions for qRT-PCR were as follows: 95°C 30s, 55°C 30s, and 72°C 90s, for a total of 40 cycles. All samples were repeated three times, and the primers used in this study are listed in Table 1.…”