Department of Neurosurgery, Inner Mongolia Baogang Hospital, Baotou City, Inner Mongolia Autonomous Region, P.R. China.
Folia Neuropathol. 2022;60(2):195-209. doi: 10.5114/fn.2022.118186.
This study was designed to elucidate the relationship of miR-211-3p and rhomboid domain containing 1 (RHBDD1) in glioma. Here, we first observed that miR-211-3p directly targets the 3˘-UTR of RHBDD1 in glioma cells using dual-luciferase reporter assay, RNA immunoprecipitation (RIP) assay, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), and Western blot analysis. Pearson's correlation analysis showed that miR-211-3p expression was negatively correlated with RHBDD1 expression in glioma tissues. CCK-8 assay, flow cytometry, and transwell assay were applied to assess cell proliferation, cell cycle distribution, migration, and invasion. The results showed that RHBDD1 knockdown inhibited cell proliferation, cell cycle G1/S transition, migration, and invasion in two glioma cell lines (U87 and LN-229). Knockdown of miR-211-3p obtained opposite results. Moreover, overexpression of RHBDD1 counteracted suppressive effects of miR-211-3p on glioma cells. Furthermore, decreased expression of CDK4, cyclin D1, N-cadherin, and vimentin as well as increased E-cadherin expression induced by miR-211-3p was reversed by RHBDD1 overexpression. Our results suggested that targeting miR-211-3p/RHBDD1 axis might be a novel effective therapeutic target for glioma treatment.
本研究旨在阐明 miR-211-3p 与脑回样结构域包含蛋白 1(RHBDD1)在胶质瘤中的关系。在这里,我们首先通过双荧光素酶报告基因检测、RNA 免疫沉淀(RIP)实验、反转录-定量聚合酶链反应(RT-qPCR)和 Western blot 分析观察到 miR-211-3p 可直接靶向胶质瘤细胞中的 RHBDD1 的 3˘-UTR。Pearson 相关性分析表明 miR-211-3p 的表达与胶质瘤组织中 RHBDD1 的表达呈负相关。CCK-8 实验、流式细胞术和 Transwell 实验用于评估细胞增殖、细胞周期分布、迁移和侵袭。结果表明,RHBDD1 敲低抑制了两种胶质瘤细胞系(U87 和 LN-229)中的细胞增殖、细胞周期 G1/S 期转变、迁移和侵袭。miR-211-3p 的敲低则获得了相反的结果。此外,RHBDD1 的过表达逆转了 miR-211-3p 对胶质瘤细胞的抑制作用。进一步研究表明,miR-211-3p 下调 CDK4、cyclin D1、N-钙黏蛋白和波形蛋白的表达并增加 E-钙黏蛋白的表达,而 RHBDD1 的过表达则逆转了这一作用。综上所述,靶向 miR-211-3p/RHBDD1 轴可能成为治疗胶质瘤的一种新的有效治疗靶点。