Abstract

Human glioma is the most common primary brain tumor and is associated with high morbidity and mortality. Aberrant expressions of microRNAs (miRNAs) are involved in glioma progression. In the present study, we aimed to elucidate the roles of miR-4530 in the pathogenesis of gliomas. miR-4530 expression was examined in human glioma clinical tissues and cell lines including U251 and T98G. The target gene of miR-4530, RTEL1, was predicted with online tools and validated by luciferase reporter assay. Lentivirus infection, transfection of plasmids, and miRNA mimics were used to manipulate gene expression. Cell proliferation was determined using the CCK-8 method, and migration and invasion assays were determined with transwell experiments. Colony formation was measured by crystal violet staining, while apoptosis was determined by Annexin V/PI staining. The anti-tumor effects of miR-4530 were evaluated in nude mice xenografted using U251 cells. Our results showed that miR-4530 was significantly down-regulated in human glioma tissues and cell lines. miR-4530 over-expression inhibited the malignant behaviors of U251 and T98G cells, including reduced proliferation, diminished colony formation, migration and invasion, and increased apoptosis. Further mechanistic investigations revealed that RTEL1 is a direct functional target of miR-4530 in gliomas, and its over-expression remarkably reverses the effects of miR-4530 mimics on inhibiting these malignant behaviors. In addition, miR-4530 over-expression inhibited the growth of xenografted U251 glioma in nude mice. Therefore, miR-4530 acts as a tumor-suppressor gene and inhibits the malignant biological behaviors of human glioma cells, which is associated with directly targeting RTEL1. The miR-4530/RTEL1 axis is a potential therapeutic target for gliomas.

中文翻译：

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miR-4530通过直接靶向RTEL1抑制人胶质瘤细胞的恶性生物学行为

摘要

人脑胶质瘤是最常见的原发性脑肿瘤，与高发病率和高死亡率有关。microRNA（miRNA）的异常表达与神经胶质瘤的进展有关。在本研究中，我们旨在阐明miR-4530在神经胶质瘤发病机理中的作用。在人神经胶质瘤临床组织和细胞系（包括U251和T98G）中检查了miR-4530的表达。miR-4530的靶基因，RTEL1，可以通过在线工具进行预测，并通过萤光素酶报告基因检测方法进行了验证。慢病毒感染，质粒转染和miRNA模拟被用来操纵基因表达。使用CCK-8方法确定细胞增殖，并通过transwell实验确定迁移和侵袭试验。通过结晶紫染色测量集落形成，而通过膜联蛋白V / PI染色确定细胞凋亡。在使用U251细胞异种移植的裸鼠中评估了miR-4530的抗肿瘤作用。我们的结果表明，miR-4530在人脑胶质瘤组织和细胞系中显着下调。miR-4530过表达抑制U251和T98G细胞的恶性行为，包括减少增殖，减少菌落形成，迁移和侵袭以及增加细胞凋亡。RTEL1是miR-4530在神经胶质瘤中的直接功能靶标，其过表达显着逆转了miR-4530模拟物抑制这些恶性行为的作用。此外，miR-4530过表达抑制裸鼠中异种移植的U251胶质瘤的生长。因此，miR-4530充当肿瘤抑制基因并抑制人类神经胶质瘤细胞的恶性生物学行为，这与直接靶向RTEL1有关。miR-4530 / RTEL1轴是神经胶质瘤的潜在治疗靶标。