【摘要】 目的探讨星形胶质细胞上调基因1（astrocyte elevated gene-1,AEG-1）表达水平对人胰腺癌细胞增殖和细胞周期的影响,研究其可能的分子机制。方法采用RNAi技术抑制As PC-1细胞中AEG-1表达,采用Western blot检测转染前后AEG-1蛋白表达情况,实验分为未转染组、脂质体组和AEG-1 si RNA转染组3组。采用MTT法和流式细胞术分析AEG-1基因沉默后对人胰腺癌As PC-1细胞生长及周期的影响。结果 Western blot检测结果显示,AEG-1 si RNA转染48 h后,As PC-1细胞中AEG-1蛋白相对灰度值为0.10±0.09,明显弱于未转染对照组的0.69±0.24和脂质体转染组的0.70±0.21（P<0.05）,而未转染对照组和脂质体转染组之间比较,差异无统计学意义（P>0.05）。转染AEG-1 si RNA的As PC-1细胞其AEG-1基因表达量下降了82%。MTT法检测结果显示,AEG-1 si RNA干扰As PC-1细胞后,AEG-1 si RNA干扰组As PC-1细胞生长减慢（P<0.05）。FCM法检测结果显示,AEG-1基因沉默后,与未转染对照组和脂质体转染组相比,G₀/G₁期细胞的比例明显升高（P<0.05）,S期的比例显著下降（均P<0.05）。结论 RNA干扰AEG-1基因表达后可明显抑制人胰腺癌As PC-1细胞增殖,且其抑制增殖作用与改变细胞周期的分布有关。更多还原

【Abstract】 Objective To explore the effect of down-regulation of astrocyte elevated gene-1（AEG-1）expression on cell proliferation and cell cycle of human pancreatic cancer cell line As PC-1,and its possible molecular mechanism.Methods The si RNA targeting at AEG-1 gene was transfected into As PC-1 cells.The protein expression of AEG-1 was detected by Western blot.The As PC-1 cells were divided into three groups:Untransfection,Lipofectamine and AEG-1si RNA-transfected group.Cell viability and cell cycle rates were determined via MTT assay and FCM.Results After 48hours’interference,the gray values of AEG-1 protein was 0.10±0.09,which was lower than in the untransfected group（0.69±0.24）as well as in the liposome transfected group（0.70±0.21,P<0.05）.However,there was no difference between untransfected group and liposome transfected group（P>0.05）.The AEG-1 expression at the gene level in AEG-1 si RNA-transfected group was reduced by 82%as compared with that in the untranfected group（P<0.05）.MTT assay showed that the proliferation of As PC-1 cells was slower than untransfected group and liposome transfected group.The percentage of cells in G₀/G₁phase in the AEG-1 Si RNA transfection group was significantly higher and the percentage of cells in S phase was significantly decreased than those in untranfected group and liposome transfected group（all P<0.05）.Conclusion The proliferation of human pancreatic cancer cell line As PC-1 cells were inhibited by AEG-1si RNA-transfection,changes in the distribution of the cell cycle was involved in the inhibiting effect when there was a lack of AEG-1 expression.更多还原