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miR-16-1 在减少神经胶质瘤细胞迁移和侵袭中发挥作用。

MiR-16-1 plays a role in reducing migration and invasion of glioma cells.

机构信息

Department of Neurosurgery, The First Affiliated Hospital of Soochow University, Suzhou, 215006, China.

出版信息

Anat Rec (Hoboken). 2013 Mar;296(3):427-32. doi: 10.1002/ar.22626. Epub 2012 Nov 23.

Abstract

MicroRNAs (miRNAs) are novel small noncoding RNA molecules that regulate gene expression at the post transcriptional level. Compelling evidence shows that there are causative links between miRNAs deregulation and cancer development and progression. This study aims to explore the functions of miR-16-1 on proliferation, apoptosis, motility, and invasion of glioma cells. Quantitative real-time PCR (qRT-PCR) was performed to detect the expression of miR-16-1 in normal brain tissues and two glioma cell lines, including U251 and U87. CCK-8, Annexin V/FITC (fluorescein isothiocyanate), wound healing, and transwell assays were used to evaluate the functions of miR-16-1 that involves cell proliferation, apoptosis, motility, and invasion. In addition, we conducted qRT-PCR to examine mRNA expression levels of Zyxin, one of putative target genes of miR-16-1, in U251 glioma cells after transfecting with miR-16-1 mimics. As a result, miR-16-1 expression level was lower in U251 and U87 cells than normal brain tissues. After miR-16-1 was upregulated in U251 cells, cellular proliferation was notably attenuated but cell apoptosis was not significantly increased. Moreover, overexpression of miR-16-1 attenuated migration and invasion of glioma cells, and U251 cells transfected with miR-16-1 showed significantly lower endogenous mRNA levels of Zyxin than those transfected with nonspecific control miRNA or mock (P < 0.05). In summary, we demonstrated that miR-16-1 expression was markedly decreased in human glioma cell lines, and for the first time, described the roles of miR-16-1 in cellular proliferation, migration, and invasion abilities of high-invasive glioma cells, and suggested that Zyxin may be one of putative target genes of miR-16-1.

摘要

微小 RNA(miRNAs)是一种新型的小非编码 RNA 分子,可在转录后水平调节基因表达。大量证据表明,miRNAs 失调与癌症的发生和发展之间存在因果关系。本研究旨在探讨 miR-16-1 对神经胶质瘤细胞增殖、凋亡、迁移和侵袭的作用。采用实时定量 PCR(qRT-PCR)检测正常脑组织和两种神经胶质瘤细胞系(U251 和 U87)中 miR-16-1 的表达。采用 CCK-8、Annexin V/FITC(荧光素异硫氰酸酯)、划痕愈合和 Transwell 实验评估 miR-16-1 涉及细胞增殖、凋亡、迁移和侵袭的功能。此外,我们还通过 qRT-PCR 检测了转染 miR-16-1 模拟物后 U251 神经胶质瘤细胞中假定的 miR-16-1 靶基因 Zyxin 的 mRNA 表达水平。结果显示,与正常脑组织相比,U251 和 U87 细胞中 miR-16-1 的表达水平较低。上调 U251 细胞中的 miR-16-1 后,细胞增殖明显减弱,但细胞凋亡无明显增加。此外,过表达 miR-16-1 可减弱神经胶质瘤细胞的迁移和侵袭,转染 miR-16-1 的 U251 细胞中 Zyxin 的内源性 mRNA 水平明显低于转染非特异性对照 miRNA 或 mock 的细胞(P<0.05)。综上所述,我们证实 miR-16-1 在人神经胶质瘤细胞系中的表达明显降低,首次描述了 miR-16-1 在高侵袭性神经胶质瘤细胞的细胞增殖、迁移和侵袭能力中的作用,并提示 Zyxin 可能是 miR-16-1 的一个潜在靶基因。

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