Chen Xiaofeng, Liu Kun, Yang Ping, Kuang Weiping, Huang Hongxing, Tu Ewen, Li Bo, Zhu Yong, Zhou Bin, Yan Lin
Department of Neurosurgery, Brain Hospital of Hunan Province, Changsha, Hunan 410007, P.R. China.
Department of Psychiatry, Brain Hospital of Hunan Province, Changsha, Hunan 410007, P.R. China.
Exp Ther Med. 2019 Jan;17(1):221-229. doi: 10.3892/etm.2018.6895. Epub 2018 Oct 29.
microRNAs (miRs) serve primary roles in certain human malignancies; however, the detailed regulatory mechanism of miR-200a in glioma progression is yet to be fully elucidated. The current study aimed to assess the expression of miR-200a in glioma as well as the regulatory mechanism of miR-200a in glioma cell proliferation, survival and invasion. RT-qPCR and western blotting were performed to examine mRNA and protein expression. An MTT assay, an EdU incorporation cell proliferation assay and a transwell assay were utilized to assess cell survival, proliferation and invasion. The results indicated that the miR-200a levels were significantly reduced in glioma tissues compared with normal brain tissues. Levels were also downregulated in glioma cell lines when compared with those in normal human astrocyte cells. Furthermore, low miR-200a expression was associated with advanced progression of glioma. The overexpression of miR-200a inhibited glioma cell proliferation, survival and invasion. Results also identified that FOXA1 was a target gene of miR-200a in glioma cells and that the increased expression of FOXA1 was negatively correlated to the decreased expression of miR-200a in glioma tissues. Furthermore, FOXA1 expression was negatively mediated by miR-200a in glioma cells and the overexpression of FOXA1 eliminated the inhibitory effects of miR-200a on the survival, proliferation and invasion of glioma cells. In conclusion, the current study demonstrated that miR-200a functions acts as a tumor suppressor in glioma by directly targeting FOXA1 and may thus be a potential candidate for the treatment of glioma.
微小RNA（miR）在某些人类恶性肿瘤中发挥着主要作用；然而，miR-200a在胶质瘤进展中的详细调控机制尚未完全阐明。本研究旨在评估miR-200a在胶质瘤中的表达以及miR-200a在胶质瘤细胞增殖、存活和侵袭中的调控机制。采用逆转录定量聚合酶链反应（RT-qPCR）和蛋白质免疫印迹法检测mRNA和蛋白质表达。利用MTT法、5-乙炔基-2'-脱氧尿苷（EdU）掺入细胞增殖试验和Transwell试验评估细胞存活、增殖和侵袭能力。结果表明，与正常脑组织相比，胶质瘤组织中miR-200a水平显著降低。与正常人星形胶质细胞相比，胶质瘤细胞系中的miR-200a水平也下调。此外，miR-200a低表达与胶质瘤的进展程度较高相关。miR-200a过表达抑制胶质瘤细胞增殖、存活和侵袭。结果还确定叉头框蛋白A1（FOXA1）是胶质瘤细胞中miR-200a的靶基因，且FOXA1表达增加与胶质瘤组织中miR-200a表达降低呈负相关。此外，miR-200a在胶质瘤细胞中负向调节FOXA1表达，FOXA1过表达消除了miR-200a对胶质瘤细胞存活、增殖和侵袭的抑制作用。总之，本研究表明miR-200a通过直接靶向FOXA1在胶质瘤中发挥肿瘤抑制作用，因此可能是治疗胶质瘤的潜在候选药物。
