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环状 RNA hsa-circ-0012129 通过靶向 microRNA-661(miR-661)促进 30 例人脑胶质瘤和人脑胶质瘤细胞系 U373、A172 和 SHG44 的细胞增殖和侵袭。

Circular RNA hsa-circ-0012129 Promotes Cell Proliferation and Invasion in 30 Cases of Human Glioma and Human Glioma Cell Lines U373, A172, and SHG44, by Targeting MicroRNA-661 (miR-661).

机构信息

Dapartment of Neurosurgery, The Third Affiliated Hospital of Bengbu Medical College, Suzhou, Anhui, China (mainland).

出版信息

Med Sci Monit. 2018 Apr 24;24:2497-2507. doi: 10.12659/msm.909229.

Abstract

BACKGROUND Circular RNA (circRNA) is a stable non-coding RNA without 5'-3' polarity and without a poly-A tail, that contains response elements for microRNAs (miRNAs) such as miR-661. There have previously been few reported studies on the role of circRNAs in glioma. The aim of this study was to investigate the effects of the expression of the circRNA, hsa-circ-0012129, and miR-661 in human glioma tissue and human glioma cell lines. MATERIAL AND METHODS Quantitative real-time polymerase chain reaction (qRT-PCR) was used to detect the expression of hsa-circ-0012129 and miR-661 in glioma tissues from 31 patients (WHO grades I-IV), compared with adjacent normal tissue, and in human glioma cell lines, U373, A172, and SHG44, compared with the normal human astrocyte cell line, NHA. The MTT assay, colony formation assay, transwell and wound scratch assays were performed to analyze and compare cell viability, cell migration, and invasion. RESULTS Expression of hsa-circ-0012129 was significantly increased in glioma tissues and cell lines; hsa-circ-0012129 knockdown significantly suppressed the proliferation, migration, and invasion abilities of U373 and SHG44 cells. A dual-luciferase reporter assay showed that hsa-circ-0012129 contained the complementary binding region with miR-661 and that hsa-circ-0012129 expression negatively regulated miR-661. Rescue experiments showed that miR-661 could reverse the effects of hsa-circ-0012129 on cell viability, cell migration and invasion of glioma cells in vitro. CONCLUSIONS The findings of this study indicated that, in human glioma cells, the circRNA, hsa-circ-0012129 might act as a natural miR-661 sponge, and that miR-661 could have suppressive effects on the expression of circ-0012129.

摘要

背景

环状 RNA(circRNA)是一种稳定的非编码 RNA,没有 5'-3'极性,也没有多聚 A 尾,其中包含 microRNAs(miRNAs)如 miR-661 的反应元件。以前关于 circRNA 在神经胶质瘤中的作用的研究很少。本研究旨在探讨 circRNA hsa-circ-0012129 和 miR-661 在人胶质瘤组织和人胶质瘤细胞系中的表达的影响。

材料和方法

采用实时定量聚合酶链反应(qRT-PCR)检测 31 例(WHO 分级 I-IV)胶质瘤组织和相邻正常组织中 hsa-circ-0012129 和 miR-661 的表达,以及人胶质瘤细胞系 U373、A172 和 SHG44 与正常人类星形胶质细胞系 NHA 相比的表达。MTT 法、集落形成实验、Transwell 和划痕实验分析比较细胞活力、细胞迁移和侵袭。

结果

hsa-circ-0012129 在胶质瘤组织和细胞系中表达明显升高;hsa-circ-0012129 敲低显著抑制 U373 和 SHG44 细胞的增殖、迁移和侵袭能力。双荧光素酶报告实验显示 hsa-circ-0012129 含有与 miR-661 互补结合的区域,hsa-circ-0012129 表达负调控 miR-661。挽救实验表明,miR-661 可以逆转 hsa-circ-0012129 对体外胶质瘤细胞活力、细胞迁移和侵袭的影响。

结论

本研究结果表明,在人胶质瘤细胞中,circRNA hsa-circ-0012129 可能作为天然 miR-661 海绵发挥作用,而 miR-661 可能对 circ-0012129 的表达具有抑制作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/33aa/5936050/c170deb8a1ce/medscimonit-24-2497-g001.jpg

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