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微小RNA-519d-3p通过靶向细胞周期蛋白D1抑制胶质瘤细胞增殖及细胞周期G1/S期转换。

MicroRNA-519d-3p inhibits cell proliferation and cell cycle G1/S transition in glioma by targeting CCND1.

作者信息

Ma Lishan, Li Jin

机构信息

Department of Neurosurgery, Wuwei People's Hospital, Wuwei, Gansu, China.

Department of Internal Neurology, Wuwei People's Hospital, Gansu, China.

出版信息

Biosci Biotechnol Biochem. 2020 Feb;84(2):297-304. doi: 10.1080/09168451.2019.1682510. Epub 2019 Oct 29.

Abstract

Glioma is the most common highly malignant primary brain tumor. MicroRNA-519d-3p exerts important effects in several tumors, but its functional role in glioma remained poorly understood. In this study, we found miR-519d-3p expression was significantly decreased in glioma tissues and cell lines. Moreover, the experiments showed that overexpression of miR-519d-3p suppressed cell proliferation and induced cell cycle G0/G1 phase arrest using MTT and flow cytometry assays in glioma cell lines, U87 and U251. Mechanistically, Cyclin D1 (CCND1) was predicted and confirmed as the direct target genes of miR-519d-3p using luciferase report assay. In addition, knockdown of CCND1 imitated the suppressive effects of miR-519d-3p on cell proliferation and cell cycle progression. Furthermore, restoration of CCND1 reversed the effects of miR-519d-3p overexpression in glioma cells. Taken together, these data demonstrate that suppression of CCND1 by miR-519d-3p might be a therapeutic target for glioma. miR-519d-3p: microRNA-519d-3p; CCND1: Cyclin D1; ATCC: American Type Culture Collection; MTT: 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide; PI: propidium iodide; WT: wild type; MUT: mutant type; SD: standard deviation.

摘要

胶质瘤是最常见的高度恶性原发性脑肿瘤。微小RNA-519d-3p在多种肿瘤中发挥重要作用,但其在胶质瘤中的功能作用仍知之甚少。在本研究中,我们发现微小RNA-519d-3p在胶质瘤组织和细胞系中的表达显著降低。此外,实验表明,在胶质瘤细胞系U87和U251中,通过MTT和流式细胞术检测,过表达微小RNA-519d-3p可抑制细胞增殖并诱导细胞周期G0/G1期阻滞。机制上,使用荧光素酶报告基因检测法预测并证实细胞周期蛋白D1(CCND1)是微小RNA-519d-3p的直接靶基因。此外,敲低CCND1可模拟微小RNA-519d-3p对细胞增殖和细胞周期进程的抑制作用。此外,恢复CCND1可逆转微小RNA-519d-3p过表达对胶质瘤细胞的影响。综上所述,这些数据表明微小RNA-519d-3p对CCND1的抑制作用可能是胶质瘤的一个治疗靶点。微小RNA-519d-3p:微小RNA-519d-3p;CCND1:细胞周期蛋白D1;ATCC:美国典型培养物保藏中心;MTT:3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐;PI:碘化丙啶;WT:野生型;MUT:突变型;SD:标准差。

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