摘要
目的探讨星形细胞上调基因1（AEG-1）表达水平对胃癌细胞增殖和细胞周期的影响，研究其可能的分子机制。方法以siRNA和AEG-1siRNA转染胃癌SGC-7901细胞后48h，将细胞分为未转染组、siRNA对照组和AEG-1siRNA转染组。应用实时荧光定量PCR和Westernblot检测转染后SGC-7901细胞中AEG-1mRNA和蛋白的表达，应用CCK-8检测细胞增殖能力，应用流式细胞术检测转染前后细胞周期的分布，应用Westernblot检测细胞增殖和细胞周期相关蛋白表达的变化。结果与未转染组和siRNA对照组比较，AEG-1siRNA转染组胃癌细胞中AEG-1蛋白的表达水平下降（P〈0．05）。与未转染组和siRNA对照组比较，AEG-1siRNA转染组AEG-1mRNA表达水平下降（P〈0．05）。与未转染组和siRNA对照组比较，AEG-1siRNA转染组转染AEG-1后的24h及其后续的各个时间点，SGC-7901细胞的增殖均明显受到抑制（均P〈0．05）。AEG-1siRNA转染组中G。／G，期细胞数比例[（61．264-1．25）％]显著高于末转染组[（46．17±1．91）％]和siRNA对照组[（46．46±1．96）％]，差异均有统计学意义（均P〈0．05）。AEG-1表达水平下降可使cdk2和cyclinD1表达水平下降以及p21表达水平升高。结论AEG-1表达水平下降抑制胃痛细胞的增殖和改变细胞周期分布．可能与cdk2、cvclinD1和D2l蛋白的表诀密切相娑．
Objective 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. Methods 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. Results 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 ceils 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 ） % ] wassignificantly higher than those in the untransfeeted 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. Conclusions 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.
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2013年第1期22-27,共6页
Chinese Journal of Oncology
基金
河南省教育厅科技攻关项目（20098320008）
河南省卫生厅科技攻关课题（200804055）
