下调Akt1表达对胆管癌HUCCA-1细胞增殖、凋亡的影响  

Down-regulating Akt1 expression influences proliferation and apoptosis of cholangiocarcinoma HUCCA-1 cells

作　　者：梁义 张小薄[2] 姚旭[2] 李跃 李殿 

机构地区：[1]辽宁省金秋医院普通外科,辽宁沈阳110001 [2]中国医科大学附属盛京医院普外科,辽宁沈阳110004

出　　处：《现代肿瘤医学》2016年第3期363-366,共4页Journal of Modern Oncology

摘　　要：目的:观察Akt1干扰后胆管癌HUCCA-1细胞增殖、细胞周期及凋亡变化,及其与下游蛋白的相关性。方法:构建干扰质粒si Akt1转染胆管癌HUCCA-1细胞,Western印迹法及qRT-PCR法检测转染效率及Akt1下调后凋亡相关基因表达。MTT法检测细胞增殖,PI法检测细胞周期,流式细胞术检测细胞凋亡。结果:MTT显示,与MOCK组和siRNA control组对比,si Akt1转染后HUCCA-1细胞增殖能力减弱,在72h及96h差异均具有显著性(P<0.05);转染si Akt1细胞停留在G1期比例增高,G2及S期比例减少(P<0.05);转染后早期凋亡及晚期凋亡率均升高(P<0.05;P<0.01)。在si Akt1转染后,凋亡相关基因p85mRNA及p-Akt1(磷酸化的Akt1)mRNA表达水平降低(P<0.05),Cleaved Caspase-9 mRNA及Cleaved Caspase-8mRNA表达升高(P<0.05)。凋亡相关基因p85及p-Akt1蛋白表达降低(P<0.05),Cleaved Caspase-9及Cleaved Caspase-8蛋白表达水平升高(P<0.05)。结论:对胆管癌HUCCA-1细胞中Akt1表达进行下调导致细胞增殖能力下降,促进细胞进入G1期,凋亡率升高,机制与其下游Cleaved Caspase基因表达具有相关性。Objective：To observe the proliferation,cell cycle and apoptosis of cholangiocarcinoma HUCCA-1 cells when Akt1 interfered and discuss the correlation with downstream proteins.Methods：Interfering plasmid siAkt1 was constructed and transfected into HUCCA-1 cells to down-regulate the expression of Akt1.Interfering effect was detected by Western blot and qRT-PCR.Proliferation,cell cycle and apoptosis were detected by MTT,PI method and flow cytometry.Results：MMT showed that compared with MOCK group and siRNA Control group,proliferation was increased in siAkt1 group at 72h and 96h（P〈0.05）.In siAkt1 group HUCCA-1 cells in G1 phase was increased,in G2 and S phases were decreased（P〈0.05）;Flow cytometry showed that the apoptosis rate in early and late phases in siAkt1 group was increased（P〈0.05,P〈0.01）.After siAkt1 transfected,gene related to apoptosis,p85 mRNA and p-Akt1 mRNA were decreased（P〈0.05）,Cleaved Caspase-9 mRNAand Cleaved Caspase-8 mRNA were increased （P〈0.05）.p85 and p-Akt1 protein was decreased（P〈0.05）,Cleaved Caspase-9 and Cleaved Caspase-8 protein was increased（P〈0.05）.Conclusion：Down-regulating Akt1 in cholangiocarcinoma HUCCA-1 cells can decrease the proliferation,inhibit promote cells staying in G1 phrase,and increase the apoptosis,the mechanism may correlate with the Cleaved Caspase gene expression.

关 键 词：胆管癌 增殖 凋亡 蛋白激酶B1 

分 类 号：R735.8[医药卫生—肿瘤]