Department of Pharmacy, Qianjiang Central Hospital of Chongqing, Chongqing, China.
Department of Pharmacy, Yongchuan Hospital of Chongqing Medical University, Chongqing, China.
J Gene Med. 2021 Jan;23(1):e3287. doi: 10.1002/jgm.3287. Epub 2020 Nov 24.
The abnormal expression of lncRNA LINC00466 (LINC00466) has been demonstrated in several tumor types. However, the expression pattern and functions of LINC00466 in glioma remain uninvestigated.
A reverse transcriptase-polymerase chain reaction (RT-PCR) was utilized to analyze LINC00466 in human glioma tissues and cell lines. Luciferase reporter assays were performed to explore whether YY1 could bind to the promoter region of LINC00466. Cell counting kit-8, flow cytometry, colony-formation, transwell migration and invasion assays were carried out to determine the involvement of INC00466 in glioma. Luciferase assays and pulldown assays were conducted to verify the binding sites.
We report that LINC00466 expression is increased in glioma cells and tissues. YY1 transcription factor (YY1) can bind directly to the LINC00466 promoter region. Clinical studies revealed that the elevated expression of LINC00466 is closely correlated with an advanced World Health Organization grade (p = 0.008), Karnofsky Performance Status score (p = 0.004) and a short overall survival (p = 0.0035) of glioma patients. Functional assays revealed that LINC00466 knockdown distinctly suppresses glioma cell proliferation, migration, invasion and epithelial-mesenchymal progress, and also promotes apoptosis. Moreover, dual-luciferase reporter assays indicated that LINC00466 acts as an endogenous sponge via binding to miR-508 and decreasing its expression. Luciferase assays and RT-PCR assays demonstrated that checkpoint kinase 1 (CHEK1) is a target of miR-508, and LINC00466 modulates CHEK1 levels by competing for miR-508. LINC00466 may exhibit its anti-oncogenic roles through targeting the miR-508/CHEK1 axis.
Our findings identified a novel glioma-related long non-coding RNA, LINC00466, which may provide a potential novel prognostic and therapeutic target for glioma.
长链非编码 RNA LINC00466（LINC00466）的异常表达已在几种肿瘤类型中得到证实。然而，LINC00466 在神经胶质瘤中的表达模式和功能仍未被研究。
利用逆转录-聚合酶链反应（RT-PCR）分析人神经胶质瘤组织和细胞系中的 LINC00466。荧光素酶报告基因检测实验用于探索 YY1 是否可以结合 LINC00466 启动子区域。细胞计数试剂盒-8、流式细胞术、集落形成、Transwell 迁移和侵袭实验用于确定 LINC00466 在神经胶质瘤中的作用。荧光素酶检测和下拉检测用于验证结合位点。
我们报告 LINC00466 在神经胶质瘤细胞和组织中表达增加。YY1 转录因子（YY1）可以直接结合 LINC00466 启动子区域。临床研究表明，LINC00466 的高表达与世界卫生组织（WHO）分级较高（p = 0.008）、卡诺夫斯基表现状态评分（KPS）较高（p = 0.004）和神经胶质瘤患者总生存期较短（p = 0.0035）密切相关。功能分析显示，LINC00466 敲低可明显抑制神经胶质瘤细胞的增殖、迁移、侵袭和上皮-间充质转化，促进细胞凋亡。此外，双荧光素酶报告基因检测实验表明，LINC00466 通过结合 miR-508 并降低其表达作为内源性海绵。荧光素酶检测和 RT-PCR 检测表明，检查点激酶 1（CHEK1）是 miR-508 的靶标，LINC00466 通过与 miR-508 竞争来调节 CHEK1 水平。LINC00466 可能通过靶向 miR-508/CHEK1 轴发挥其抗癌作用。
我们的研究结果确定了一种新的神经胶质瘤相关长链非编码 RNA，LINC00466，它可能为神经胶质瘤提供一种新的潜在预后和治疗靶点。
