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微小RNA-182通过靶向F-box蛋白7促进神经胶质瘤进展。

MiR-182 promotes glioma progression by targeting FBXW7.

作者信息

Liu Shiming, Liu Hanbo, Deng Min, Wang Haowen

机构信息

Department of Interventional Medicine, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, PR China.

Department of Interventional Medicine, Zhejiang Provincial People's Hospital, People's Hospital of Hangzhou Medical College, PR China.

出版信息

J Neurol Sci. 2020 Apr 15;411:116689. doi: 10.1016/j.jns.2020.116689. Epub 2020 Jan 16.

Abstract

OBJECTIVE

MicroRNAs (miRNAs) are widely considered to play an important role in the tumor progression. In this study, we aimed to investigate the potential biological effects of miR-182 and its target FBXW7 on glioma development.

METHODS

Expression data of glioma were procured from TCGA database. Differential analysis was performed to identify the potential differentially expressed miRNA (DEmiRNA), and bioinformatics databases were utilized for target genes prediction. qRT-PCR was conducted to detect miR-182 and western blot was performed to test FBXW7 in protein level. Then the cells were processed for proliferation determination by CCK-8, migration test through wound healing assay, invasion detection via Transwell and apoptosis measurement by flow cytometry. In addition, dual-luciferase reporter gene assay was carried out for evaluation of the targeted relationship between the miR-182 and FBXW7.

RESULTS

MiR-182 was found to be up-regulated in glioma cells while FBXW7 was down-regulated. Besides, miR-182 was predicted to targeted bind with FBXW7 on 3'UTR via bioinformatics methods, and their targeted relationship was further validated by dual-luciferase assay. MiR-182 mimic could significantly promote cell proliferation, migration, invasion and inhibit cell apoptosis, while miR-182 inhibitor had the negative effect. Moreover, FBXW7 knockdown could reverse the inhibitory effect of miR-182 inhibitor on glioma cells.

CONCLUSION

MiR-182 promotes cell proliferation, migration, invasion and inhibits cell apoptosis in glioma by targeting FBXW7.

摘要

目的

微小RNA(miRNA)被广泛认为在肿瘤进展中起重要作用。在本研究中,我们旨在探讨miR-182及其靶标FBXW7对神经胶质瘤发展的潜在生物学效应。

方法

从TCGA数据库获取神经胶质瘤的表达数据。进行差异分析以鉴定潜在的差异表达miRNA(DEmiRNA),并利用生物信息学数据库预测靶基因。进行qRT-PCR检测miR-182,进行蛋白质印迹检测FBXW7的蛋白水平。然后通过CCK-8检测细胞增殖,通过伤口愈合试验进行迁移试验,通过Transwell检测侵袭,并通过流式细胞术测量凋亡。此外,进行双荧光素酶报告基因测定以评估miR-与FBXW7之间的靶向关系。

结果

发现miR-182在神经胶质瘤细胞中上调,而FBXW7下调。此外,通过生物信息学方法预测miR-182在3'UTR上与FBXW7靶向结合,并且通过双荧光素酶测定进一步验证了它们的靶向关系。miR-182模拟物可显著促进细胞增殖、迁移、侵袭并抑制细胞凋亡,而miR-182抑制剂则具有相反作用。此外,FBXW7基因敲低可逆转miR-182抑制剂对神经胶质瘤细胞的抑制作用。

结论

miR-182通过靶向FBXW7促进神经胶质瘤细胞的增殖、迁移、侵袭并抑制细胞凋亡。

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