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Circ_0080425通过miR-1182/FGF9轴影响高糖环境下血管内皮细胞功能

Circ_0080425 affects the function of endothelial cells in hyperglycemic environment via miR-1182/FGF9 axis

    摘要
  • 摘要:
    目的 探讨circ_0080425调控高糖环境下内皮功能的作用及分子机制。
    方法 采用qRT-PCR检测在高糖环境下培养的人血管内皮细胞(human vascular endothelial cells，HUVECs)中circ_0080425的表达变化，通过小干扰RNA（siRNA）对circ_0080425进行敲低，并利用CCK-8、平板克隆、Transwell和流式细胞术，观察circ_0080425对高糖培养下的HUVECs增殖、迁移以及凋亡等生物学特性的影响。利用生物信息学网站预测circ_0080425的下游miRNA以及靶基因，并通过双荧光素酶报告实验验证分子间的相互结合。此外，通过sponge慢病毒以及质粒转染对目标分子的表达水平进行调控，并采用CCK-8、平板克隆、Transwell和流式细胞术等进行功能学验证。
    结果 Circ_0080425在高糖培养条件下HUVECs中表达上调，且circ_0080425可通过miR-1182/FGF9轴调控高糖条件下HUVECs的增殖、迁移和凋亡（P＜0.05）。
    结论 Circ_0080425可通过miR-1182/FGF9轴影响高糖环境下的血管内皮功能。

     

    Abstract:
    Objective To explore the role and molecular mechanism of circ_0080425 in regulating the function of endothelial cells in hyperglycemic environment.
    Methods The expression difference of circ_0080425 in human vascular endothelial cells (HUVECs) cultured with medium containing high glucose or normal medium was confirmed by qRT-PCR. Circ_0080425 was knockdown by siRNA transfection. The effects of circ_0080425 on the proliferation, migration and apoptosis of HUVECs under high glucose condition were investigated using CCK-8 assay, colony formation, Transwell and Flow cytometry. The Circinteractome was used to predict the downstream miRNAs of circ_0080425 and the TargetScanHuman 8.0 tool was used to predict the target genes of the miRNAs. The intermolecular interactions were verified by dual luciferase reporter assay. In addition, the target molecules were regulated by sponge virus as well as plasmid transfection, and the function was verified by CCK-8 assay, colony formation, Transwell and Flow cytometry.
    Results The expression of circ_0080425 was up-regulated in HUVECs under the condition of high glucose. Circ_0080425 can promote the proliferation, migration, and inhibit apoptosis of HUVECs under high glucose condition through miR-1182/FGF9 axis (P < 0.05).
    Conclusions  Circ_0080425 regulates the function of endothelial cells in hyperglycemic environment via miR-1182/FGF9 axis.

     

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