Ma Lishan, Li Jin
Department of Neurosurgery, Wuwei People's Hospital, Wuwei, Gansu, China.
Department of Internal Neurology, Wuwei People's Hospital, Gansu, China.
Biosci Biotechnol Biochem. 2020 Feb;84(2):297-304. doi: 10.1080/09168451.2019.1682510. Epub 2019 Oct 29.
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.
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。