siRNA沉默蛋白激酶Cε基因抑制胆管细胞癌生长及其作用机制  

Protein kinase C-epsilon gene silenced by siR NA inhibits the development of cholangiocellular carcinoma and its mechanism

作　　者：郭明明[1] 刘江辉[1] 蔡锐彬[1] 王晶[2] 陈斌[3] 

机构地区：[1]中山大学附属第一医院急诊科,广州510080 [2]中山大学附属第一医院妇产科,广州510080 [3]中山大学附属第一医院肝胆外科,广州510080

出　　处：《中华肝脏外科手术学电子杂志》2014年第6期44-47,共4页Chinese Journal of Hepatic Surgery(Electronic Edition)

基　　金：国家自然科学基金(81302550)

摘　　要：目的探讨小干扰RNA(si RNA)沉默蛋白激酶Cε(PKCε)基因抑制胆管细胞癌的生长及其作用机制。方法分别将PKCε-si RNA及阴性对照(NC)-si RNA转染至人胆管细胞癌QBC939细胞中,设立PKC组和NC组。另设未转染对照组(CTRL组)。采用细胞计数试剂盒(CCK)-8法检测细胞增殖抑制率,流式细胞术检测细胞凋亡率,蛋白质印迹法检测PKCε、survivin蛋白表达情况。3组比较采用单因素方差分析,两两比较采用LSD-t检验。结果 PKC组细胞24、48、72 h的细胞增殖抑制率逐渐增高,分别为(7.52±0.33)%、(15.28±0.20)%和(37.12±0.45)%,与CTRL组比较差异有统计学意义(LSD-t=15.37,27.12,35.05;P<0.05)。PKC组细胞凋亡率(56.9±6.1)%明显高于CTRL组的(12.5±1.3)%(LSD-t=28.55,P<0.05)。PKC组PKCε、survivin蛋白表达水平较NC组、CTRL组降低。结论 si RNA沉默PKCε基因可能通过下调survivin蛋白表达,促进细胞凋亡从而抑制胆管细胞癌的生长。ObjectiveTo investigate the role of protein kinase C-epsilon （PKCε） gene silenced by small interfering ribonucleic acid （siRNA） in inhibiting the development of cholangiocellular carcinoma and its mechanism.MethodsHuman cholangiocellular carcinoma QBC939 cells were respectively transfected using PKCε-siRNA and negative control （NC）-siRNA to establish PKC group and NC group. And untransfected control （CTRL） group was established. Cell counting kit （CCK）-8 assay was used to define the cell proliferation inhibition rate. Cell apoptosis rate was detected by flow cytometery. The expressions of protein PKCε and survivin were detected by Western blot. The comparison of three groups was conducted using one way analysis of variance and pairwise comparison using LSD-t test.ResultsThe cell proliferation inhibition rates at 24, 48, 72 h [（7.52±0.33）%, （15.28±0.20）%, （37.12±0.45）% ] increased gradually in PKC group, where significant difference was observed compared with those in CTRL group （LSD-t=15.37, 27.12, 35.05;P〈0.05）. The cell apoptosis rate was （56.9±6.1）% in PKC group, which was significantly higher compared with that in CTRL group＆nbsp;[（12.5±1.3） % ] （LSD-t=28.55,P〈0.05）. The expressions of protein PKCε and survivin decreased in PKC group compared with those in NC group and CTRL group.ConclusionPKCε gene silenced by siRNA may inhibit the development of cholangiocellular carcinoma through down-regulating the expression of protein survivin and promoting cell apoptosis.

关 键 词：胆管肿瘤 RNA干扰 蛋白激酶CΕ 细胞增殖 细胞凋亡 

分 类 号：R735.8[医药卫生—肿瘤]