Effect of 14-3-3β gene silenced by small interfering RNA on biological behavior of glioma cells and its mechanism
10.3760/cma.j.issn.1671-8925.2014.09.003
- VernacularTitle:RNA干预沉默14-3-3β基因对胶质瘤细胞生物学行为的影响及其机制
- Author:
Fanghe GONG
1
;
Jing YE
;
Tiandong LI
;
Hongmin BAI
;
Shuai LIU
;
Weimin WANG
;
Guoliang WANG
Author Information
1. 广州军区广州总医院神经医学专科医院
- Keywords:
Glioma cell;
14-3-3β gene;
RNA interfere;
Cell invasion Cell proliferation;
Cell apoptosis
- From:
Chinese Journal of Neuromedicine
2014;13(9):876-880
- CountryChina
- Language:Chinese
-
Abstract:
Objective To explore the effect of 14-3-3 β gene on biological behavior ofglioma cell line and its mechanism.Methods Conventional cultured SVGp12,U251,U87 and SHG-44 cell lines and U251 cells silenced by 14-3-3[β-small interfering RNA (siRNA) were collected; real time-PCR and Western blotting were used to detect the 14-3-3β gene and protein expressions in these cells.Conventional cultured U251 cells at logarithmic phase were divided into three groups:experimental group (14-3-3β-siRNA transfection),negative control group (siRNA transfection) and blank control group; 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay was used to assess the proliferation of U251 cells,flow cytometry was used to test the cell apoptosis,and cell migration was analyzed by Transwell chamber assay.Results As compared with those in the normal glial cells,14-3-3β gene and protein expression levels in the glioma cells were significantly higher (P＜0.05); as compared with negative control and blank control groups,U251 cells in the experimental group had significantly decreased gene and protein expressions of 14-3-3β,decreased proliferation and migration abilities,significantly increased apoptosis rate and p53 mRNA level (P＜0.05).Conclusion Silence of 14-3-3 β gene decreases U251 cells proliferation and migration through p53 mediated pathway; consequently,a new explanation about how 14-3-3 β regulates glioma cells proliferation and migration can be clarified,and a potential target for glioma treatment can be provided.
