Let-7i regulates glioma U251 stem cell differentiation through LIN28 protein
10.3760/cma.j.issn.1671-8925.2014.09.002
- VernacularTitle:let-7i通过调节LIN28影响胶质瘤U251干细胞成熟分化
- Author:
Dongyi JIANG
1
;
Lei SHI
;
Yi WAN
;
Xifeng FEI
;
Zhimin WANG
Author Information
1. 上海交通大学附属苏州九龙医院神经外科
- Keywords:
Let-7i;
Glioma;
Stem cells;
LIN28
- From:
Chinese Journal of Neuromedicine
2014;13(9):870-875
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the effect of MicroRNA (miRNA) let-7i on differentiation ofglioma U251 stem cells and the potential mechanisms.Methods Glioma U251 cells were cultured in vitro and CD133 positive cells were sorted by magnetic cell separation.Composite let-7i mimics and let-7i controls were transfected into U251 cells; the let-7i expression was detected by real time-PCR; the expressions ofCD133,nestin and LIN28 were detected by Western blotting; the differentiation status was assessed via glial fibrillary acidic protein (GFAP) immunocytochemistry.Composite LIN28 siRNA and siRNA control were transfected into U251 cells; the expressions of CD133 and nestin were detected by Western blotting.Targetscan software analysis and luciferase reporter system were employed to verified the possibility of LIN28 being the target gene of let-7i.Results let-7i in the U251 cells were over-expressed in group of let-7i mimics,enjoying 17.9 fold of group of let-7i control; the expressions of CD 133,nestin and LIN28 were 13.9％,43.7％ and 53.6％,separately,of group of let-7i control; GFAP positive labeling index in group oflet-7i mimics (83.0±1.93)％ was significantly higher than that of group oflet-7i control (39.7±6.73)％ (P＜0.05).The expressions of CD133 and nestin in group of LIN28 siRNA decreased to 23.7％ and 37.9％ of those in groups of siRNA control.LIN28 3'UTR existed let-7i paired binding sites and LIN28 could be the target gene of let-7i.Conclusion over-expressing let-7i could significantly down-regulate the expressions of CD133 and nestin,and enhance the glioma stem cell differentiation through the potential mechanism of down-regulating the expression of LIN28.
