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Effect of silencing AEG-1 with small interfering RNA on the proliferation and cell cycle of gastric carcinoma SGC-7901 cells / 中华肿瘤杂志
Chinese Journal of Oncology ; (12): 22-27, 2013.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-284246
Responsible library: WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To 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.</p><p><b>METHODS</b>Control 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.</p><p><b>RESULTS</b>Real-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.</p><p><b>CONCLUSIONS</b>The 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.</p>
Subject(s)
Full text: Available Database: WPRIM (Western Pacific) Main subject: Pathology / Stomach Neoplasms / RNA, Messenger / Transfection / Cell Adhesion Molecules / Down-Regulation / Cyclin D1 / RNA, Small Interfering / RNA Interference / Cell Line, Tumor Limits: Animals / Female / Humans Language: Chinese Journal: Chinese Journal of Oncology Year: 2013 Document type: Article
Full text: Available Database: WPRIM (Western Pacific) Main subject: Pathology / Stomach Neoplasms / RNA, Messenger / Transfection / Cell Adhesion Molecules / Down-Regulation / Cyclin D1 / RNA, Small Interfering / RNA Interference / Cell Line, Tumor Limits: Animals / Female / Humans Language: Chinese Journal: Chinese Journal of Oncology Year: 2013 Document type: Article
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