Li Xiaojuan, Cao Quansheng, Wang Yanyu, Wang Yongsheng
Department of Central Sterile Supply, The First Affiliated Hospital of Henan University of Science and Technology No. 24, Jinghua Road Luoyang 471003 China
Department of Cardiovascular Surgery, The First Affiliated Hospital of Henan University of Science and Technology Luoyang China.
RSC Adv. 2019 Dec 17;9(71):41709-41719. doi: 10.1039/c9ra08322g. eCollection 2019 Dec 13.
Long non-coding RNA OIP5-AS1 (lncRNA OIP5-AS1) and microRNA-128-3p (miRNA-128-3p) have been reported to play significant roles in human diseases. However, their role in atherosclerosis (AS) has been less studied. The aim of this research was to reveal the roles and functional mechanisms of OIP5-AS1 and miRNA-128-3p in AS development. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot assays were performed to detect gene expression. Cell proliferation and apoptosis were assessed using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry analysis, respectively. In addition, ELISA was employed to determine the levels of interleukin (IL)-6, IL-1β, and tumor necrosis factor (TNF)-α. Oxidative stress was examined using a relevant kit. Furthermore, the interaction between miR-128-3p and OIP5-AS1 or cyclin-dependent kinase inhibitor 2A (CDKN2A) was predicted using StarBase, and then confirmed using the dual-luciferase reporter assay or the RNA immunoprecipitation (RIP) assay. We found that OIP5-AS1 and CDKN2A levels were upregulated and the miR-128-3p level was downregulated in oxidized low-density lipoprotein (ox-LDL)-induced THP1 cells. OIP5-AS1 knockdown weakened the regulatory effect of ox-LDL on cell progression. Interestingly, OIP5-AS1 directly interacted with miR-128-3p and miR-128-3p-targeted CDKN2A. Furthermore, OIP5-AS1 regulated ox-LDL-induced cell progression through modulating miR-128-3p expression, and miR-128-3p exerted its influence by modulating the CDKN2A level. Finally, we confirmed that OIP5-AS1 suppressed miR-128-3p expression to increase the level of CDKN2A. In conclusion, our findings demonstrate that OIP5-AS1 knockdown repressed the effect of ox-LDL on cell progression through regulating the miR-128-3p/CDKN2A axis, providing a potential target for the treatment of AS.
据报道，长链非编码RNA OIP5-AS1（lncRNA OIP5-AS1）和微小RNA-128-3p（miRNA-128-3p）在人类疾病中发挥重要作用。然而，它们在动脉粥样硬化（AS）中的作用研究较少。本研究的目的是揭示OIP5-AS1和miRNA-128-3p在AS发展中的作用及其功能机制。采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法检测基因表达。分别使用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐（MTT）法和流式细胞术分析评估细胞增殖和凋亡。此外，采用酶联免疫吸附测定（ELISA）法测定白细胞介素（IL）-6、IL-1β和肿瘤坏死因子（TNF）-α的水平。使用相关试剂盒检测氧化应激。此外，利用StarBase预测miR-128-3p与OIP5-AS1或细胞周期蛋白依赖性激酶抑制剂2A（CDKN2A）之间的相互作用，然后通过双荧光素酶报告基因测定法或RNA免疫沉淀（RIP）测定法进行验证。我们发现，在氧化型低密度脂蛋白（ox-LDL）诱导的THP1细胞中，OIP5-AS1和CDKN2A水平上调，而miR-128-3p水平下调。敲低OIP5-AS1可减弱ox-LDL对细胞进程的调节作用。有趣的是，OIP5-AS1直接与miR-128-3p相互作用，且miR-128-3p靶向CDKN2A。此外，OIP5-AS1通过调节miR-128-3p的表达来调控ox-LDL诱导的细胞进程，而miR-128-3p则通过调节CDKN2A水平发挥作用。最后，我们证实OIP5-AS1抑制miR-128-3p的表达以增加CDKN2A的水平。总之，我们的研究结果表明，敲低OIP5-AS1可通过调节miR-128-3p/CDKN2A轴来抑制ox-LDL对细胞进程的影响，为AS的治疗提供了一个潜在靶点。
