RNA干扰沉默水通道蛋白3基因对人肝癌细胞SMMC-7721增殖和凋亡的影响  

Effect of Aquaporin 3 Gene Silencing on the Proliferation and Apoptosis of the Hepatocellular Carcinoma Cell Line SMMC-7721

作　　者：付雪岩 王雅玮[2] 王文青 吴刚[1] FU Xueyan;WANG Yawei;WANG Wenqing;WU Gang(Department of General Surgery,The First Hospital,China Medical University,Shenyang 110001,China;Department of Geriatric Surgery,The First Hospital,China Medical University,Shenyang 110001,China)

机构地区：[1]中国医科大学附属第一医院普通外科,沈阳110001 [2]中国医科大学附属第一医院老年外科,沈阳110001

出　　处：《中国医科大学学报》2019年第7期596-600,共5页Journal of China Medical University

基　　金：辽宁省自然科学基金(2015020529)

摘　　要：目的研究沉默人肝癌细胞SMMC-7721的水通道蛋白3(AQP3)基因表达对细胞增殖和凋亡的影响。方法体外培养人肝癌细胞SMMC-7721细胞;siRNA-AQP3沉默AQP3基因表达;实时PCR和Westernblotting检测干扰后AQP3mRNA和蛋白的表达水平;CCK8和平板克隆检测细胞增殖能力;AnnexinVFITC/PI双标记法检测细胞凋亡率;研究设空白对照组(Neg.cont)、siRNA-NC组、siRNA-AQP3#1组、siRNA-AQP3#2组。结果siRNA沉默AQP3基因后,AQP3mRNA和蛋白的表达水平明显降低(P<0.001,P<0.01)。在siRNA-AQP3#1组和siRNA-AQP3#2组中,CCK8实验显示其细胞活力明显降低;平板克隆也得到相似的结果(P<0.01,P<0.05)。流式细胞仪检测结果显示,siRNA-AQP3#1组和siRNA-AQP3#2组细胞凋亡比率明显高于对照组(P<0.001,P<0.01)。结论RNA干扰沉默AQP3基因能够抑制人肝癌细胞SMMC-7721细胞增殖,促进细胞凋亡。Objective To investigate the effect of Aquaporin 3 (AQP3) gene silencing on the proliferation and apoptosis of the hepatocellular carcinoma cell line,SMMC-7721.Methods Two kinds of small interfering RNA( siRNA) were separately transfected into SMMC-7721 cells using Lipofectamine 2000.The silencing efficiency was evaluated using qRT-PCR and Western blotting.SMMC-7721 cell proliferation was determined using CCK-8 and colony-forming assays.Moreover,SMMC-7721 cell apoptosis was measured using the Annexin V-FITC/PI double staining method.The experiment was performed with negative control( NC),siRNA-NC,siRNA-AQP3#1,and siRNA-AQP3#2 groups.Results AQP3 mRNA and protein expression levels( P < 0.001 and P < 0.01,respectively) were significantly decreased after siRNA-mediated AQP3 gene silencing.Moreover,CCK-8 and colony-forming assays showed a significant decrease in cell viability( P < 0.01,P < 0.05) in the siRNA-AQP3#1 and siRNA-AQP3#2 groups.The flow cytometry-determined apoptosis rates of the siRNA-AQP3#1 and siRNA-AQP3#2 groups were significantly higher than that of the control group( P < 0.001,P < 0.01).Conclusion RNA silencing of AQP3 gene can inhibit SMMC-7721 cell proliferation and promote cell apoptosis.

关 键 词：肝细胞癌 SMMC-7721 水通道蛋白3 RNA干扰 增殖 凋亡 

分 类 号：R735.7[医药卫生—肿瘤]