沉默ABCC4基因表达对人胰腺癌PANC1、BxPC-3细胞增殖的影响  

Effect of ABCCA gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANCl and BxPC-3

作　　者：马瑾[1] 张卓[2] 王建承[2] 

机构地区：[1]上海交通大学医学院附属瑞金医院卢湾分院消化内科,上海200020 [2]上海交通大学医学院附属瑞金医院普外科

出　　处：《中华胰腺病杂志》2013年第4期252-255,共4页Chinese Journal of Pancreatology

摘　　要：目的检测ABCCA基因沉默后对人胰腺癌PANC1、BxPC-3细胞增殖及细胞周期的影响。方法构建插入靶向ABCCA的shRNA片段的重组慢病毒载体，感染人胰腺癌细胞株PANCI和BxPC-3。采用实时定量PCR法和蛋白质印迹法检测感染细胞的ABCCAmRNA及蛋白表达，克隆形成实验测定感染细胞形成的克隆数，流式细胞仪检测感染细胞的细胞周期。结果成功构建了插入靶向ABCCA的shRNA片段的重组慢病毒载体。重组慢病毒感染PANC1及BxPC-3细胞后，细胞ABCCAmRNA表达被抑制（0．28±0．01比1．00±0．03，0．22±0．02比1．00±0．03，P值均〈0．05）；ABCCA蛋白表达亦显著下调；感染的PANCl细胞克隆形成数量显著减少（4比65，P〈0．05）；细胞周期阻滞在G，期[（54．98±1．78）％比（42．93±0．88）％，（68．55±0．75）％比（54．76±0．29）％]。结论沉默人胰腺癌PANCl和BxPC-3细胞的ABCC4基因表达可显著抑制癌细胞的增殖，使细胞阻滞于G1期。Objective To determine the effect of ABCC4 gene silencing on cell proliferation and cell cycle in human pancreatic cancer cell lines PANC1 and BxPC-3. Methods PANC1 and BxPC-3 pancreatic cancer cells were transfected with a lentivirus expressing an ABCCA short hairpin RNA （shRNA）. ABCC4 mRNA and protein expression of transfeeted cells was determined by RT-PCR and Western blot, colony formation ability was measured by colony assay, and cell cycle change was investigated by the flow eytometrie analysis. Results Lentivirns expressing an ABCCA short hairpin RNA （shRNA） was successfully established. After transfection with shRNA lentivirus, ABCC4 mRNA and protein expression were significantly inhibited （0.28 ± 0. 01 vs 1. 00 ± 0.03,0.22 ±0. 02 vs 1.00 ±0. 03, P 〈 0.05 ）. Colony formation ability was significantly decreased （4 vs 65, P 〈0.05） , and cell cycle was significantly blocked at G1 phase [ （54.98 ± 1.78） % vs （ 42.93 ± 0.88 ） %, （ 68.55 ± 0.75 ） % vs （ 54.76 ± 0.29 ） % ]. Conclusions ABCCA gene silencing can significantly inhibit cell proliferation of human pancreatic cancer cell lines PANC1 and BxPC-3, and block the cells at G1 phase.

关 键 词：胰腺肿瘤 ABCC4 细胞增殖 RNA干扰 慢病毒感染 

分 类 号：R735.9[医药卫生—肿瘤]