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目的:探讨长链非编码RNA CRNDE对胶质瘤干细胞特性改变的影响及其机制。方法:应用免疫磁珠法从人胶质瘤细胞系U251中分离出CD133n +U251干细胞(U251s)。将实验细胞分为U251s组、U251s-CRNDE组及U251组、U251-CRNDE组，其中U251s-CRNDE组、U251-CRNDE组细胞预先用CRNDE短发夹RNA(shRNA)慢病毒载体转染以下调CRNDE的表达。应用实时荧光定量PCR(RT-qPCR)检测各组细胞中n CRNDE mRNA的表达水平，应用细胞计数试剂(CCK)-8法检测各组细胞的增殖变化，应用Transwell小室实验检测各组细胞的侵袭能力，应用划痕实验检测各组细胞的迁移能力，应用平板克隆实验检测各组细胞的克隆形成能力，应用流式细胞术检测各组细胞的周期改变，应用Western blotting法检测U251s组、U251s-CRNDE组细胞中干细胞标志蛋白A2B5、CD133、巢蛋白、Sox2、Oct-4、Nanog以及磷脂酰肌醇激酶(PI3K)-蛋白激酶B(AKT)-雷帕霉素靶蛋白(mTOR)信号通路关键蛋白PI3K、AKT、磷酸化(p)-AKT、mTOR蛋白的表达水平。n 结果:与U251组相比，U251s组细胞中n CRNDE mRNA的表达水平明显升高，差异有统计学意义(n P<0.05)。U251s-CRNDE组与U251s组细胞相比，U251-CRNDE组与U251组细胞相比，前者的细胞增殖率、穿膜细胞数量、细胞迁移率、细胞集落形成数目、G2/M期细胞比例明显降低，G1/G0期细胞比例明显升高，差异均有统计学意义(n P<0.05)。与U251s组相比，U251s-CRNDE组细胞中CD133、巢蛋白、Sox2、Oct-4及PI3K、AKT、p-AKT、mTOR蛋白的表达水平明显降低，差异均有统计学意义(n P<0.05)。n 结论:CRNDE在胶质瘤干细胞中表达增高，而下调CRNDE的表达后可以抑制胶质瘤干细胞的特性，且这种影响与PI3K-AKT-mTOR信号通路的调控有关。“,”Objective:To investigate the influence of long non-coding RNA-CRNDE in stem cell properties and their mechanism in glioma.Methods:The CD133n +U251 stem cells (U251s) were isolated from human glioma cell line U251 by immunomagnetic beads. Cells were divided into U251s group, U251s-CRNDE group, U251 group, and U251-CRNDE group; CRNDE shRNA lentiviral vectors were transfected into cells in the U251s-CRNDE group and U251-CRNDE group to down-regulate the CRNDE expression. The n CRNDE mRNA expression was detected by real-time fluorescent quantitative PCR (RT-qPCR); CCK-8 assay was used to detect the cell viability; Transwell assay was used to detect the cell invasion; wound healing method was used to detect the cell migration; plate cloning assay was employed to detect the clone formation; flow cytometry was used to detect the cell cycles; Western blotting was used to detect the expressions of A2B5, CD133, nestin, Sox2, Oct-4 and Nanog, as well as phosphatidylinositol kinase (PI3K)-protein kinase B (AKT)-mammalian target of rapamycin (mTOR) signal pathway key proteins (PI3K, AKT, phosphorylated-AKT, and mTOR).n Results:The n CRNDE mRNA expression in U251s group was significantly higher than that in U251 group (n P<0.05). As compared with U251s group, U251s-CRNDE group had significantly decreased cell viability, cell invasion and cell migration, statistically smaller number of cell colony, significantly decreased proportion of cells in G2/M phase, significantly increased proportion of cells in G1/G0 phase (n P<0.05); and the same trend was noted between U251 group and U251-CRNDE group. As compared with U251s group, U251s-CRNDE group had significantly decreased expressions of CD133, nestin, Sox2, Oct-4, PI3K, AKT, phosphorylated-AKT, and mTOR (n P<0.05).n Conclusion:The CRNDE expression is increased in glioma stem cells, and down-regulation of CRNDE expression can inhibit the differentiation, metabolism and proliferation of glioma stem cells; and this effect is related to the regulation of PI3K-AKT-mTOR signaling pathway.