siRNA干扰AEG-1对人胰腺癌AsPC-1细胞增殖和细胞周期的影响  

Effect of AEG-1 siRNA on proliferation and cell cycle distribution of human pancreatic cancer AsPC-1 cells

作　　者：黄炎[1] 干耘[1] 钱春艳[2] 徐新伟[1] 戴丹 戚显忠 朱莉[1] 谌伽莉 董水凤 任国平[3] HUANG Yan;GAN Yun;QIAN Chun-yan(Department of Pathology,the First People＇ s Hospital of Yuhang District,Hangzhou,Zhejiang 311100,China)

机构地区：[1]杭州市余杭区第一人民医院病理科,浙江杭州311100 [2]杭州市余杭区第一人民医院检验科 [3]浙江大学附属第一医院病理科,浙江杭州310000

出　　处：《中华全科医学》2018年第10期1618-1622,共5页Chinese Journal of General Practice

基　　金：浙江省医药卫生一般研究计划(2013KYB228);杭州市余杭区重大科研项目(余科[2012]68号2012-5医学)

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,G_0/G_1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。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.

关 键 词：胰腺癌 RNA干扰 星形胶质细胞上调基因1 细胞周期 

分 类 号：R735.9[医药卫生—肿瘤] R329.28[医药卫生—临床医学]