CAJ | 학술논문

目的探讨小干扰RNA（siRNA）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-siRNA及阴性对照（NC）-siRNA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 siRNA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。
목적탐토소간우RNA（siRNA）침묵단백격매Cε(PKCε)기인억제담관세포암적생장급기작용궤제。방법분별장PKCε-siRNA급음성대조（NC）-siRNA전염지인담관세포암QBC939세포중，설립PKC조화NC조。령설미전염대조조（CTRL조）。채용세포계수시제합（CCK）-8법검측세포증식억제솔，류식세포술검측세포조망솔，단백질인적법검측PKCε、survivin단백표체정황。3조비교채용단인소방차분석，량량비교채용LSD-t검험。결과PKC조세포24、48、72 h적세포증식억제솔축점증고，분별위（7.52±0.33）%、（15.28±0.20）%화（37.12±0.45）%，여CTRL조비교차이유통계학의의(LSD-t =15.37，27.12，35.05；P<0.05)。PKC조세포조망솔（56.9±6.1）%명현고우CTRL조적（12.5±1.3）%(LSD-t=28.55，P<0.05)。PKC조PKCε、survivin단백표체수평교NC조、CTRL조강저。결론 siRNA침묵PKCε기인가능통과하조survivin단백표체，촉진세포조망종이억제담관세포암적생장。
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 [(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.