Qi Zengxin, Cai Shengyong, Cai Jiajun, Chen Lingchao, Yao Yu, Chen Liang, Mao Ying
Department of Neurosurgery, Huashan Hospital, Fudan University, 12 Wulumuqi Middle Road, Shanghai, 200040, China.
Department of Neurosurgery, Shanghai Medical College, Fudan University, Shanghai, China.
BMC Neurol. 2016 Dec 1;16(1):248. doi: 10.1186/s12883-016-0769-y.
MicroRNAs are significantly involved in tumorigenesis and progression of glioma. However, the critical part they play in glioma have not been fully elaborated. miR-491 and Tripartite motif containing 28 (TRIM28) are reported to aberrantly express in glioblastoma multiforme (GBM). Here, we detected miR-491 and TRIM28 expression and function in glioma cells.
We analyzed miR-491 expressions in 20 primary human GBM tissues and 6 control brain tissues by qRT-PCR assays and searched for The Cancer Genome Atlas (TCGA) database. Then we predicted possible mRNA target of miR-491 by TargetScan/MicroRNA and confirmed it via luciferase reporter assays. Knock-down of miR-491 and transfection of pLenti-TRIM28 were performed in U251 and U87 cells. Proliferation ability was examined by MTT and clone formation assays.
miR-491 expression was obviously reduced in GBM cells and tissues. There was a positive correlation between the down-regulation of miR-491 and poor prognosis. Spearman's correlation analysis demonstrated that miR-491 expression was negatively correlated with TRIM28 protein level. Possible mRNA binding sites of miR-491 predicted by TargetScan/MicroRNA were proved by luciferase assays. Clone formation and MTT assays indicated that up-regulation of miR-491 inhibited the proliferation of glioma cells.
miR-491 regulates glioma cells proliferation in vitro by targeting TRIM28.
微小RNA在胶质瘤的发生和发展中发挥着重要作用。然而，它们在胶质瘤中所起的关键作用尚未得到充分阐明。据报道，miR-491和含三联基序蛋白28（TRIM28）在多形性胶质母细胞瘤（GBM）中异常表达。在此，我们检测了miR-491和TRIM28在胶质瘤细胞中的表达及功能。
我们通过qRT-PCR分析了20例原发性人类GBM组织和6例对照脑组织中miR-491的表达，并检索了癌症基因组图谱（TCGA）数据库。然后，我们通过TargetScan/MicroRNA预测了miR-491可能的mRNA靶点，并通过荧光素酶报告基因检测进行了验证。在U251和U87细胞中进行miR-491敲低和pLenti-TRIM28转染。通过MTT和克隆形成试验检测增殖能力。
GBM细胞和组织中miR-491表达明显降低。miR-491下调与预后不良呈正相关。Spearman相关性分析表明，miR-491表达与TRIM28蛋白水平呈负相关。TargetScan/MicroRNA预测的miR-491可能的mRNA结合位点经荧光素酶试验证实。克隆形成和MTT试验表明，miR-491上调抑制了胶质瘤细胞的增殖。
miR-491通过靶向TRIM28在体外调节胶质瘤细胞增殖。
