长链非编码 RNA GHET1 通过下调 miR-216a 促进胶质瘤细胞系 U251 的活力、迁移和侵袭。
Long non-coding RNA GHET1 promotes viability, migration and invasion of glioma cell line U251 by down-regulation of miR-216a.
机构信息
Department of Neurosurgery, Jining No. 1 People's Hospital, Jining, Shandong, China.
出版信息
Eur Rev Med Pharmacol Sci. 2019 Feb;23(4):1591-1599. doi: 10.26355/eurrev_201902_17118.
OBJECTIVE
Glioma is among the most aggressive of all human malignancies. Long non-coding RNA (lncRNA) gastric carcinoma highly expressed transcript 1 (GHET1) was considered an important oncogene in tumors. However, the role of GHET1 in glioma was rarely studied. The present work aimed to explore the effect of GHET1 on glioma cell line U251.
MATERIALS AND METHODS
The viability, migration and invasion of U251 cells were analyzed by Cell Counting Kit-8 (CCK-8) assay, and transwell migration/invasion assay, respectively. The relative expression of GHET1 and miR-216a were detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). The expression of cell cycle-related proteins, cell metastasis-associated proteins and main factors in the JAK2/STAT3 and p53/survivin pathways was analyzed by Western blot. Cell transfection assay was conducted to alter expression of GHET1 and miR-216a in U251 cells.
RESULTS
The viability, migration and invasion of U251 cells were promoted by GHET1 overexpression. The pro-cell cycle genes including Cyclin D1, CDK4 and CDK6, and the pro-metastasis genes including MMP-9 and Vimentin were up-regulated after GHET1 was overexpressed. MiR-216a was found to be down-regulated by GHET1 overexpression, and it was involved in the effects of GHET1 on U251 cells. GHET1 might promote U251 cells by down-regulating miR-216a. Finally, we found that GHET1 overexpression activated the JAK2/STAT3 and p53/survivin signaling pathways by down-regulating miR-216a.
CONCLUSIONS
GHET1 overexpression increased viability, migration and invasion of U251 cells by down-regulating miR-216a. Mechanically, GHET1-miR-216a axis activated the JAK2/STAT3 and p53/survivin signaling pathways.
目的
神经胶质瘤是人类所有恶性肿瘤中侵袭性最强的肿瘤之一。长链非编码 RNA（lncRNA）胃癌高表达转录本 1（GHET1）被认为是肿瘤中的重要癌基因。然而，GHET1 在神经胶质瘤中的作用很少被研究。本研究旨在探讨 GHET1 对神经胶质瘤细胞系 U251 的影响。
材料和方法
通过细胞计数试剂盒-8（CCK-8）检测和 Transwell 迁移/侵袭检测，分别分析 U251 细胞的活力、迁移和侵袭。通过定量实时聚合酶链反应（qRT-PCR）检测 GHET1 和 miR-216a 的相对表达。通过 Western blot 分析细胞周期相关蛋白、细胞转移相关蛋白以及 JAK2/STAT3 和 p53/survivin 通路中的主要因子的表达。通过细胞转染实验改变 U251 细胞中 GHET1 和 miR-216a 的表达。
结果
GHET1 过表达促进 U251 细胞的活力、迁移和侵袭。过表达 GHET1 后，细胞周期相关基因包括 Cyclin D1、CDK4 和 CDK6 以及转移相关基因 MMP-9 和 Vimentin 上调。发现 GHET1 过表达下调 miR-216a，miR-216a 参与 GHET1 对 U251 细胞的作用。GHET1 可能通过下调 miR-216a 促进 U251 细胞。最后，我们发现 GHET1 过表达通过下调 miR-216a 激活 JAK2/STAT3 和 p53/survivin 信号通路。
结论
GHET1 过表达通过下调 miR-216a 增加 U251 细胞的活力、迁移和侵袭。机制上，GHET1-miR-216a 轴激活 JAK2/STAT3 和 p53/survivin 信号通路。
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