RNAi技术沉默DKK1基因对食管癌ECA109及EC1细胞系增殖的影响

目的:通过构建DKK1的靶向小干扰RNA(small interfering RNA),探讨干扰DKK1基因的表达和对食管癌ECA109及EC1细胞系增殖的影响.方法:转染DKK1 siRNA于食管癌细胞系ECA109、EC1,应用蛋白免疫印迹方法检测转染前后细胞中DKK1的蛋白表达变化.食管癌细胞系ECA109及EC1成功干扰DKK1表达后,应用CCK-8法检测癌细胞增殖变化,平板克隆法检测癌细胞的克隆形成能力变化.结果:食管癌细胞系转染DKK1 siRNA后,ECA109中DKK1蛋白表达降低48.62％ (P ＜0.01),EC1中DKK1蛋白表达降低50％(P＜0.01).在CCK-8法检测癌细胞增殖变化实验中,DKK1 siRNA转染后24 h、48 h及72 h,相比阴性对照组,细胞增殖能力显著降低.平板克隆实验中,DKK1 siRNA转染后,ECA109细胞克隆均数(77.53±4.948)低于空白对照组(44.2±7.704),克隆率下降42.98％,而EC1细胞克隆均数(71.67±5.239)低于空白对照组(36±2.646),克隆率下降49.76％.结论:干扰DKK1的表达能够抑制食管癌细胞的增殖,DKK1可能成为抑制食管癌增殖的分子靶点.

Effect of silencing DKK1 gene on esophageal cancer on the proliferation of ECA109 and EC1 cell lines by RNA interference

Objective:To investigate the effect of DKK1 siRNA on the proliferation of ECA109 and EC1 cell lines by constructing small interfering RNA of DKK1.Methods:DKK1 siRNA was transfected into esophageal cancer cell lines ECA109 and EC1,and the protein expression of DKK1 was detected by immunoblatting method.After ECM109 and EC1 were successfully transfected into the esophageal cancer cell lines,the proliferation of cancer cells was detected by CCK-8 method.The clonogenic ability of cancer cells was detected by plate cloning method.Results:DKK1 protein was down-regulated by 48.62％ (P ＜0.01) and decreased by 50％ (P ＜0.01) in DKK1 siRNA transfected ECM109 and EC1 cells.The proliferation of DKK1 siRNA-treated ECM109 and EC1 cells was significantly decreased at 24 h,48 h and 72 h in CCK-8 assay compared with the negative control group.The clonal number of ECA109 and EC1 cells in DKK1 siRNA-treated group was significantly lower than that in control group,and the clone rate of ECA109 and EC1 decreased by 42.98％ and 49.76％.Conclusion:Knock-down of DKK1 contributed to inhibit the proliferation of esophageal cancer cells,and DKK1 might be a molecular target to inhibit the proliferation of esophageal carcinoma.