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长链非编码 RNA NEAT1 通过 miR-361-5p/VEGFA 通路促进血管瘤的进展。

Long non-coding RNA NEAT1 promotes the progression of hemangioma via the miR-361-5p/VEGFA pathway.

机构信息

Department of Aesthetic Plastic & Craniofacial Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.

Department of Aesthetic Plastic & Craniofacial Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.

出版信息

Biochem Biophys Res Commun. 2019 May 14;512(4):825-831. doi: 10.1016/j.bbrc.2019.03.084. Epub 2019 Mar 27.

Abstract

Hemangioma (HA) is the most common benign vascular neoplasm of infancy that is resulted from abnormal proliferation of endothelial cells. Recent studies demonstrated that long non-coding RNAs (lncRNAs) were closely related to the pathogenesis of HA. LncRNA Nuclear enriched abundant transcript 1 (NEAT1) was involved in multiple tumor formation and biological behaviors of endothelial cells. However, the role and molecular mechanism of NEAT1 in HA are still unknown. The expression levels of NEAT1 and miR-361-5p were detected in proliferating phase HAs, involuting phase HAs, and normal skin tissues. The role and mechanism of NEAT1 on the proliferation, migration and apoptosis of hemangioma endothelial cells (HemECs) were analyzed by Cell Counting Kit (CCK)-8, transwell, flow cytometry, caspase-3 activity, dual-luciferase assay, RNA immunoprecipitation, Biotin-labeled miR-361-5p pulldown assay and western blot by gain- and loss-of-functions. We found that compared with normal skin tissues, NEAT1 expression was elevated, whereas miR-361-5p decreased in HA tissues especially in proliferating phase HAs. Downregulation of NEAT1 significantly suppressed the viability, PCNA expression and migration, but increased apoptotic cell numbers and caspase-3 activity of HemECs. NEAT1 functioned as a competing endogenous RNA to regulate VEGFA expression via sponging miR-361-5p. Taken together, these findings indicate that NEAT1 promotes the proliferation and migration, whereas inhibits the apoptosis of HemECs via regulating miR-361-5p/VEGFA axis.

摘要

血管瘤(HA)是最常见的婴儿期良性血管肿瘤,是由内皮细胞异常增殖引起的。最近的研究表明,长链非编码 RNA(lncRNA)与 HA 的发病机制密切相关。lncRNA 核丰富丰富转录物 1(NEAT1)参与了多种肿瘤的形成和内皮细胞的生物学行为。然而,NEAT1 在 HA 中的作用和分子机制尚不清楚。检测增殖期 HA、消退期 HA 和正常皮肤组织中 NEAT1 和 miR-361-5p 的表达水平。通过细胞计数试剂盒(CCK)-8、Transwell、流式细胞术、caspase-3 活性、双荧光素酶报告基因检测、RNA 免疫沉淀、生物素标记的 miR-361-5p 下拉实验和 Western blot 分析 NEAT1 对血管内皮细胞(HemECs)增殖、迁移和凋亡的作用和机制。通过 gain- 和 loss-of-functions 实验。我们发现,与正常皮肤组织相比,HA 组织中 NEAT1 表达升高,而 miR-361-5p 表达降低,尤其是在增殖期 HA 组织中。下调 NEAT1 可显著抑制 HemECs 的活力、PCNA 表达和迁移,但增加凋亡细胞数量和 caspase-3 活性。NEAT1 通过海绵 miR-361-5p 作为竞争内源性 RNA 调节 VEGFA 表达。总之,这些发现表明,NEAT1 通过调节 miR-361-5p/VEGFA 轴促进 HemECs 的增殖和迁移,同时抑制其凋亡。

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