Influence of long non-coding RNA-CRNDE in stem cell properties and their mechanism in glioma
10.3760/cma.j.cn115354-20191216-00750
- VernacularTitle: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
- Author:
Genghuan WANG
1
;
Heping SHEN
;
Huan HUANG
;
Xiaohong JIANG
;
Peng CHEN
Author Information
1. 嘉兴学院附属第二医院神经外科 314000
- Keywords:
Glioma;
Long non-coding RNA;
CRNDE;
Tumor stem cell;
Stem cell property
- From:
Chinese Journal of Neuromedicine
2021;20(2):133-140
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the influence of long non-coding RNA-CRNDE in stem cell properties and their mechanism in glioma.Methods:The CD133 +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 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). Results:The CRNDE mRNA expression in U251s group was significantly higher than that in U251 group ( 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 ( 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 ( P<0.05). 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.
