Jiang Yu, Wang Xiaohui, Zhang Ji, Lai Renchun
State Key Laboratory of Oncology in South China, Department of Anesthesiology, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong 510060, P.R. China.
State Key Laboratory of Oncology in South China, Department of Radiation Oncology, Collaborative Innovation Center for Cancer Medicine, Sun Yat-sen University Cancer Center, Guangzhou, Guangdong 510060, P.R. China.
Oncol Lett. 2018 Jul;16(1):1243-1252. doi: 10.3892/ol.2018.8727. Epub 2018 May 16.
MicroRNAs (miRNAs/miRs) serve tumor promoting or suppressive roles in different human cancer types, including glioma; however, the regulatory underlying mechanism by which miR-599 affects glioma progression remains largely unknown. The aim of the present study was to investigate the expression of miR-599 in glioma, as well as the underlying regulatory mechanism. Reverse transcription-quantitative polymerase chain reaction and western blot analysis were used to examine mRNA and protein expression, respectively. MTT, wound healing and transwell assays were conducted to study cell proliferation, migration and invasion, respectively. A dual-luciferase reporter gene assay was used to confirm the targeting association between miR-599 and Ras-related protein Rab-27B (hereafter RAB27B). In the present study, miR-599 expression was observed to be significantly downregulated in human glioma tissues and cell lines, when compared with normal brain tissues and normal human astrocyte cells, respectively. Low miR-599 expression was significantly associated with glioma progression. Ectopic expression of miR-599 caused a significant reduction in the proliferation, migration and invasion of U-87MG Uppsala and U251 cells. Bioinformatics analysis and dual-luciferase reporter gene assay data identified that RAB27B was a direct target gene of miR-599. The mRNA and protein expression of RAB27B was significantly downregulated following miR-599 overexpression in U-87MG Uppsala and U251 cells. Rescue experiments demonstrated that RAB27B rescued the miR-599-induced inhibition of glioma cell growth, migration and invasion. In addition, RAB27B expression was significantly upregulated in glioma tissues and cell lines, and the expression levels of RAB27B were inversely correlated with miR-599 levels in glioma tissues; therefore, the present study demonstrated that miR-599 exerts a tumor-suppressive role in glioma progression by targeting RAB27B, indicating that miR-599 may be used as a potential candidate for glioma treatment.
微小RNA(miRNA/miR)在包括神经胶质瘤在内的不同人类癌症类型中发挥着促肿瘤或抑肿瘤作用;然而,miR-599影响神经胶质瘤进展的潜在调控机制仍 largely未知。本研究的目的是调查miR-599在神经胶质瘤中的表达及其潜在调控机制。分别采用逆转录-定量聚合酶链反应和蛋白质印迹分析来检测mRNA和蛋白质表达。分别进行MTT、伤口愈合和Transwell实验来研究细胞增殖、迁移和侵袭。采用双荧光素酶报告基因实验来确认miR-599与Ras相关蛋白Rab-27B(以下简称RAB27B)之间的靶向关联。在本研究中,与正常脑组织和正常人星形胶质细胞相比,分别观察到miR-599在人类神经胶质瘤组织和细胞系中的表达显著下调。低miR-599表达与神经胶质瘤进展显著相关。miR-599的异位表达导致U-87MG Uppsala和U251细胞的增殖、迁移和侵袭显著降低。生物信息学分析和双荧光素酶报告基因实验数据确定RAB27B是miR-599的直接靶基因。在U-87MG Uppsala和U251细胞中,miR-599过表达后,RAB27B的mRNA和蛋白质表达显著下调。拯救实验表明,RAB27B挽救了miR-599诱导的神经胶质瘤细胞生长、迁移和侵袭的抑制作用。此外,RAB27B在神经胶质瘤组织和细胞系中的表达显著上调,且神经胶质瘤组织中RAB27B的表达水平与miR-599水平呈负相关;因此,本研究表明miR-599通过靶向RAB27B在神经胶质瘤进展中发挥抑肿瘤作用,表明miR-599可能作为神经胶质瘤治疗的潜在候选物。