Effect of silencing AEG-1 with small interfering RNA on the proliferation and cell cycle of gastric carcinoma SGC-7901 cells.
- VernacularTitle:siRNA下调星形细胞上调基因1表达对胃癌SGC-7901细胞增殖和细胞周期的影响
- Author:
Cai-feng ZHANG
1
;
Yong-hua XIA
;
Qing-fen ZHENG
;
Zhen-juan LI
;
Xiao-he GUO
;
Hui-cong ZHOU
;
Li-li ZHANG
;
Liang-peng DONG
;
Yu HAN
;
Zhu-e LIU
;
Wen-ju WANG
;
Yan-li LUO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Adhesion Molecules; biosynthesis; genetics; Cell Cycle Checkpoints; Cell Line, Tumor; Cell Proliferation; Cyclin D1; metabolism; Cyclin-Dependent Kinase 2; metabolism; Cyclin-Dependent Kinase Inhibitor p21; metabolism; Down-Regulation; Female; Humans; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; RNA Interference; RNA, Messenger; metabolism; RNA, Small Interfering; genetics; Stomach Neoplasms; metabolism; pathology; Transfection
- From: Chinese Journal of Oncology 2013;35(1):22-27
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo explore the effect of down-regulation of astrocyte elevated gene-1 (AEG-1) expression on cell proliferation and cell cycle of gastric carcinoma cells, and its possible molecular mechanism.
METHODSControl siRNA and AEG-1 siRNA were transfected into gastric carcinoma SGC-7901 cells. 48 h after transfection, the cells were divided into 3 groups including untransfected, siRNA control and AEG-1 siRNA transfection groups. Expressions of AEG-1 mRNA and protein in the 3 group cells were detected by real-time quantitative PCR and Western blot. The changes of cell proliferation were examined using CCK-8 kit, and the cell cycle distribution was detected by flow cytometry. Finally, expressions of cell proliferation and cell cycle related proteins were detected by Western blot.
RESULTSReal-time quantitative PCR and Western blot demonstrated that compared with the untransfected and siRNA control groups, expressions of AEG-1 mRNA and protein were significantly down-regulated in the AEG-1 siRNA transfection group (P < 0.05), but there was no significant difference between the untransfected and siRNA control groups (P > 0.05). Furthermore, in vivo experiment confirmed a significant down-regulation of AEG-1 protein in the AEG-1 siRNA transfection group (P < 0.05). In addition, AEG-1 siRNA obviously inhibited the proliferation of SGC-7901 cells at different time points after transfection with AEG-1 siRNA. The percentage of cells in G0/G1 phase in the AEG-1 siRNA transfection group [(61.26 ± 1.25)%] was significantly higher than those in the untransfected group [(46.17 ± 1.91)%] and siRNA control group [(46.46 ± 1.96)%], and there was a significant difference between them (all P < 0.001). Furthermore, the result of Western blotting revealed that down-regulation of AEG-1 expression evoked the down-regulation of cdk2 and cyclin D1 expressions and elevation of p21 expression in vitro and in vivo.
CONCLUSIONSThe inhibition of cell proliferation and cell cycle arrest mediated by down-regulation of AEG-1 expression may be closely associated with the changes of expression of cell cycle related proteins including cdk2, cyclin D1 and p21.
