Department of Neurosurgery, Nanjing Medical University Affiliated Changzhou No. 2 People's Hospital, Changzhou, 213003, Jiangsu, China.
Department of Neurology, Xuzhou Third People's Hospital, Xuzhou, Jiangsu, China.
Hum Cell. 2020 Apr;33(2):416-426. doi: 10.1007/s13577-020-00331-z. Epub 2020 Feb 18.
Glioma is one of the most common malignant tumors and shows a high metastasis rate and poor prognosis. Abnormal expression of long non-coding RNAs (lncRNAs) contributes to various human tumors including gliomas. This study aimed to investigate the regulatory role of the antisense RNA of growth arrest special 5 (GAS5-AS1), a novel lncRNA, in gliomas. Expression of GAS5-AS1 and microRNA-106b-5p (miR-106b-5p) in glioma tissues and cells was detected by quantitative reverse transcription PCR, northern blotting, or fluorescent in situ hybridization. Cell proliferation, migration, and invasion were analyzed by CCK-8 and Transwell assays. BALB/c nude mice were used to establish a glioma xenograft animal model by subcutaneous injection of U251 cells transfected with small interfering RNA targeting GAS5-AS1. A dual-luciferase reporter assay was conducted to confirm the targeting relationship between GAS5-AS1 and miR-106b-5p. GAS5-AS1 expression was downregulated in glioma tissues and cells, and upregulation of GAS5-AS1 expression inhibited glioma cell proliferation, migration, and invasion. GAS5-AS1 expression was correlated with the glioma tumor grade. In nude mice, upregulation of GAS5-AS1 markedly suppressed glioma tumor growth. GAS5-AS1 overexpression significantly increased the miR-106b-5p level in glioma cells, and GAS5-AS1 expression was negatively related to miR-106b-5p expression in glioma tissues. Overexpression of miR-106b-5p reversed the inhibitory effects of GAS5-AS1 upregulation on glioma cell proliferation and metastasis, while restoration of TUSC2 rescued the proliferation and invasion of glioma cells transfected with miR-106b-5p mimics. In summary, lncRNA GAS5-AS1 inhibited glioma proliferation, migration, and invasion by sponging miR-106b-5p and regulating the expression of TUSC2. Our results suggest GAS5-AS1 as a novel target for the treatment and prognosis prediction of gliomas.
神经胶质瘤是最常见的恶性肿瘤之一，具有较高的转移率和较差的预后。长链非编码 RNA（lncRNA）的异常表达与包括神经胶质瘤在内的各种人类肿瘤有关。本研究旨在探讨生长停滞特异基因 5（GAS5-AS1）反义 RNA 的调节作用，这是一种新的 lncRNA，在神经胶质瘤中的作用。通过定量逆转录 PCR、 northern 印迹或荧光原位杂交检测神经胶质瘤组织和细胞中 GAS5-AS1 和 microRNA-106b-5p（miR-106b-5p）的表达。通过 CCK-8 和 Transwell 测定分析细胞增殖、迁移和侵袭。通过皮下注射转染 GAS5-AS1 小干扰 RNA 的 U251 细胞建立神经胶质瘤异种移植动物模型。通过双荧光素酶报告实验证实 GAS5-AS1 与 miR-106b-5p 之间的靶向关系。在神经胶质瘤组织和细胞中下调 GAS5-AS1 的表达，上调 GAS5-AS1 的表达抑制神经胶质瘤细胞的增殖、迁移和侵袭。GAS5-AS1 的表达与神经胶质瘤肿瘤分级相关。在裸鼠中，上调 GAS5-AS1 显著抑制神经胶质瘤肿瘤生长。GAS5-AS1 过表达显著增加神经胶质瘤细胞中的 miR-106b-5p 水平，GAS5-AS1 的表达与神经胶质瘤组织中 miR-106b-5p 的表达呈负相关。过表达 miR-106b-5p 逆转了 GAS5-AS1 上调对神经胶质瘤细胞增殖和转移的抑制作用，而恢复 TUSC2 挽救了转染 miR-106b-5p 模拟物的神经胶质瘤细胞的增殖和侵袭。总之，lncRNA GAS5-AS1 通过海绵 miR-106b-5p 抑制神经胶质瘤的增殖、迁移和侵袭，并调节 TUSC2 的表达。我们的研究结果表明，GAS5-AS1 是治疗和预测神经胶质瘤的新靶点。
