Background

Glioma has the chief type of primary brain tumors worldwide. The glioma may be controlled by regulators including some lncRNAs, miRNAs, and proteins.

Objective

Our study aims to discover the underlying mechanism for lncPCAT19/miR-142-5p/MELK axis in glioma progression.

Methods

The clinical samples were from patients with gliomas in our Hospital. Hematoxylin–eosin staining (H&E) was applied to determine the clinical pathological changes. Real time PCR was performed to measure the levels of lncPCAT19, miR-142-5p, MELK, and expression of other genes. Western blot was conducted to detect the protein level of MELK. RIP assay was performed to analyze the interaction between lncPCAT19 and miR-142-5p, and dual-luciferase reporter assay was used to determine the binding site between lncPCAT19 and miR-142-5p. CCK-8, colony formation assay, flow cytometry, and trans-well assay were carried out to confirm cell proliferation, colony formation, apoptosis, and invasion, respectively.

Results

LncPCAT19 was increased in cancer tissues. Then, lncPCAT19 could interact with and down-regulate miR-142-5p. Knockdown of lncPCAT19 distinctly inhibited tumor growth in vivo. Interfering lncPCAT19/overexpression of miR-142-5p decreased glioma cell proliferation, colony formation and invasion, and promoted cell apoptosis by down-regulating expression of Cyclin B1, CDK2, N-cadherin, Bcl-2, and by up-regulating expression of Bax and E-cadherin. Moreover, overexpression of lncPCAT19 overturned tumor-suppressing role of miR-142-5p in cells. Additionally, lncPCAT19 and miR-142-5p synergistically regulated expression of MELK. In conclusion, lncPCAT19 enhanced glioma development via increasing MELK by performing as a sponge of miR-142-5p.

Conclusions

LncPCAT19 promotes glioma progression by sponging miR-142-5p to upregulate MELK levels. Thus, lncPCAT19/miR-142-5p/MELK signaling would be a potential target for glioma treatment.

中文翻译：

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抑制长链非编码 RNA PCAT19 抑制胶质瘤细胞增殖和侵袭，并通过调节 miR-142-5p 靶向的 MELK 增加细胞凋亡

背景

胶质瘤是全世界原发性脑肿瘤的主要类型。神经胶质瘤可能受调节剂控制，包括一些 lncRNA、miRNA 和蛋白质。

客观的

我们的研究旨在发现 lncPCAT19/miR-142-5p/MELK 轴在胶质瘤进展中的潜在机制。

方法

临床样本来自我院胶质瘤患者。苏木精-伊红染色（H＆E）用于确定临床病理变化。进行实时 PCR 以测量 lncPCAT19、miR-142-5p、MELK 的水平和其他基因的表达。进行蛋白质印迹以检测MELK的蛋白质水平。进行RIP分析以分析lncPCAT19与miR-142-5p之间的相互作用，并使用双荧光素酶报告基因分析确定lncPCAT19与miR-142-5p之间的结合位点。分别进行 CCK-8、集落形成测定、流式细胞术和跨孔测定以确认细胞增殖、集落形成、凋亡和侵袭。

结果

LncPCAT19 在癌组织中增加。然后，lncPCAT19 可以与 miR-142-5p 相互作用并下调。敲除 lncPCAT19 明显抑制体内肿瘤生长。干扰lncPCAT19/miR-142-5p的过表达通过下调细胞周期蛋白B1、CDK2、N-钙粘蛋白、Bcl-2的表达，上调神经胶质瘤细胞的增殖、集落形成和侵袭，促进细胞凋亡。 Bax 和 E-钙粘蛋白。此外，lncPCAT19 的过表达推翻了 miR-142-5p 在细胞中的肿瘤抑制作用。此外，lncPCAT19 和 miR-142-5p 协同调节 MELK 的表达。总之，lncPCAT19 作为 miR-142-5p 的海绵，通过增加 MELK 来增强神经胶质瘤的发展。

结论

LncPCAT19 通过海绵 miR-142-5p 上调 MELK 水平来促进胶质瘤进展。因此，lncPCAT19/miR-142-5p/MELK 信号将成为神经胶质瘤治疗的潜在靶点。