Department of Neurosurgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China.
CNS Neurosci Ther. 2019 Aug;25(8):842-854. doi: 10.1111/cns.13121. Epub 2019 Mar 28.
In this study, we examined the expression of lncRNA ENST00000413528 in glioma and determined its role in glioma development.
LncRNA ENST00000413528 was detected in glioma tissues by lncRNA microarray. Then, we performed real-time PCR, CCK-8, colony formation assay, flow cytometry, caspase-3/7 assay and animal experiment to detect the function of ENST00000413528 in glioma after ENST00000413528 knockdown. Subsequent bioinformatics analysis, luciferase reporter assays and RNA immunoprecipitation (RIP) assay western blotting indicated possible downstream regulatory molecules. The expression of PLK1 in glioma tissues was also examined by immunohistochemistry staining.
Expression of ENST00000413528 was significantly increased in glioma tissues and LN229 and U251 cells. PLK1 protein could not be detected in peritumoral brain edema (PTBE) tissues; however, it showed an increasing number of positively cytoplasmic stained from WHO-Grade II to Grade III gliomas. Knockdown of ENST00000413528 in glioma cells inhibited cell proliferation and colony formation abilities, induced the G0/G1 arrest of the cell cycle, and promoted apoptosis. The dual reporter assay and RNA immunoprecipitation assay verified the interaction between ENST00000413528 and miR-593. We also demonstrated that polo-like kinase 1 (PLK1) was regulated by miR-593; PLK1 messenger RNA lacking 3'UTR partially reversed the effects caused by ENST00000413528 knockdown or miR-593 upregulation.
lncRNA ENST00000413528 is closely related to the development of glioma via the miR-593-5p/PLK1 pathway.
本研究旨在检测长链非编码 RNA ENST00000413528 在脑胶质瘤中的表达，并确定其在脑胶质瘤发生发展中的作用。
采用长链非编码 RNA 芯片检测脑胶质瘤组织中 lncRNA ENST00000413528 的表达情况。然后，通过实时 PCR、CCK-8 法、集落形成实验、流式细胞术、caspase-3/7 检测和动物实验，检测 ENST00000413528 敲低后对脑胶质瘤的作用。随后的生物信息学分析、荧光素酶报告基因实验和 RNA 免疫沉淀（RIP）实验及 Western blot 检测表明可能存在下游调控分子。采用免疫组化染色检测脑胶质瘤组织中 PLK1 的表达。
ENST00000413528 在脑胶质瘤组织和 LN229、U251 细胞中的表达明显升高。PLK1 蛋白在瘤周水肿（PTBE）组织中无法检测到，但从 WHO 分级 II 级到 III 级脑胶质瘤中，其细胞质阳性染色数量逐渐增多。脑胶质瘤细胞中 ENST00000413528 敲低可抑制细胞增殖和集落形成能力，诱导细胞周期 G0/G1 期阻滞，并促进细胞凋亡。双报告基因检测和 RNA 免疫沉淀实验验证了 ENST00000413528 与 miR-593 的相互作用。我们还证明了 polo 样激酶 1（PLK1）受 miR-593 调控；缺少 3'UTR 的 PLK1 信使 RNA 部分逆转了 ENST00000413528 敲低或 miR-593 上调引起的作用。
lncRNA ENST00000413528 通过 miR-593-5p/PLK1 通路与脑胶质瘤的发生发展密切相关。
