Inhibitory effect of Notch2 gene transfection on growth of glioma cell line A172 in vitro
10.3760/cma.j.issn.1671-8925.2014.02.001
- VernacularTitle:转染Notch2基因对人脑胶质瘤细胞系A172生长的抑制作用
- Author:
Guangxiu WANG
1
;
Peng XU
;
Zhifan JIA
;
Anling ZHANG
;
Peiyu PU
Author Information
1. 天津医科大学总医院神经病学研究所神经肿瘤实验室
- Keywords:
Notch2;
Glioma;
Proliferation;
Apoptosis;
EGFR/PI3K/AKT pathway
- From:
Chinese Journal of Neuromedicine
2014;13(2):109-113
- CountryChina
- Language:Chinese
-
Abstract:
Objective To investigate the inhibitory effect of Notch2 gene on the growth of A172 glioblastoma cells and its underlying mechanism.Methods Blank control group,empty vector transfection group (pcDNA3) and Notch2 plasmid transfection group (Notch2-pcDNA3) were employed in our study; A172 glioma cells in the later two groups were transfected with blank load plasmid and Notch2 plasmid,respectively.Cell growth was detected by MTT assay,and cell cycle and apoptosis were analyzed by flow cytometry.Transwell assay was used to study the cell invasion,and the expressions of important signaling pathway members,including Notch2,epidermal growth factor receptor (EGFR),phospho-Akt (P-Akt),phosphatidylinositol 3-kinase (PI3K),phosphatase and tensin homologue deleted on chromosome 10 (PTEN),caspase3,nuclear factor κB (NF-κB),proliferating cell nuclear antigen (PCNA),intedeukin-2 (Bcl-2),matrix metalloproteinase-2 (MMP-2),MMP-9andcyclin-dependent kinase inhibitor 1 (Cyclin D1),were detected by Western blotting.Results MTT assay showed that the cell viability in the A172 cells of Notch2-pcDNA3 group was significantly decreased as compared with that in the blank control cell and pcDNA3 groups (P＜0.05); decreased cell number at S phase and G2/M phase was noted in A172 cells of Notch2-pcDNA3 group,while cell number at G0/G1 phase increased by 22.1％ as compared with that in the blank control group; the percentage of apoptotic cells in the A172 cells of Notch2-pcDNA3 group (14.8％) was significantly increased as compared with that in the blank control cells (1.2％); the number of invaded cells in the A172 cells of Notch2-pcDNA3 group (17.43) was significantly decreased as compared with that in the blank control cells (35.23).The important members related to PI3K-AKT signaling pathway,including EGFR,p-AKT,PI3K,NF-κB,PCNA,Bcl-2,MMP-2,MMP-9 and Cyclin D1 had obvious decreased expression,while PTEN and Caspase-3,the antagonistic PI3K/AKT activity tumor suppressor proteins,had obvious increased expression.Conclusion The suppressive effect of Notch2 gene on glioma cell proliferation may result from the inhibition of EGFR/PI3K/AKT pathway.
