Department of Neurosurgery, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.
Department of Operation, the First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, 710061, China.
Acta Biochim Biophys Sin (Shanghai). 2020 Dec 29;52(12):1394-1403. doi: 10.1093/abbs/gmaa126.
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.
人类脑胶质瘤是最常见的原发性脑肿瘤，其发病率和死亡率都很高。微小 RNA（miRNA）的异常表达参与了胶质瘤的进展。在本研究中，我们旨在阐明 miR-4530 在胶质瘤发病机制中的作用。检测了 miR-4530 在人类脑胶质瘤临床组织和细胞系（包括 U251 和 T98G）中的表达。使用在线工具预测 miR-4530 的靶基因 RTEL1，并通过荧光素酶报告基因实验进行验证。慢病毒感染、质粒转染和 miRNA 模拟物用于操纵基因表达。使用 CCK-8 法测定细胞增殖，用 Transwell 实验测定迁移和侵袭实验。用结晶紫染色法测定集落形成，用 Annexin V/PI 染色法测定细胞凋亡。通过 U251 细胞的裸鼠异种移植评估 miR-4530 的抗肿瘤作用。我们的结果表明，miR-4530 在人类脑胶质瘤组织和细胞系中显著下调。miR-4530 的过表达抑制了 U251 和 T98G 细胞的恶性行为，包括增殖减少、集落形成减少、迁移和侵袭减少以及凋亡增加。进一步的机制研究表明，RTEL1 是胶质瘤中 miR-4530 的直接功能靶基因，其过表达显著逆转了 miR-4530 模拟物对抑制这些恶性行为的作用。此外，miR-4530 的过表达抑制了裸鼠异种移植 U251 脑胶质瘤的生长。因此，miR-4530 作为一种肿瘤抑制基因，抑制了人类脑胶质瘤细胞的恶性生物学行为，其作用与直接靶向 RTEL1 有关。miR-4530/RTEL1 轴是胶质瘤的潜在治疗靶点。
