目的:以shRNA抑制食管癌ECA109细胞内Survivin的表达,探讨Survivin shRNA干扰对CDK4基因表达的影响。方法:ECA109细胞分为Survivin shRNA干扰组、质粒对照组和空白细胞组,采用RNA干扰技术沉默ECA109细胞株中Survivin基因的表达,RT-PCR检测各组细胞中Survivin和CDK4 mRNA的表达,Western blot方法检测各组细胞Survivin蛋白的表达,流式细胞术检测细胞周期的改变。结果:与质粒对照组及空白细胞组比较,Survivin shRNA干扰组细胞Survivin mRNA及蛋白表达下调,CDK4 mRNA表达明显降低,差异均具有统计学意义(P均〈0.05);Survivin shRNA干扰组G2期延长,G2期细胞比例增加,细胞G2期增殖阻滞。结论:Survivin基因可能在转录水平对CDK4具有调控作用,其具体调控机制有待进一步研究。
To study Survivin shRNA interference on CDK4 mRNA and protein levels of esophageal cells ECA109. METHODS:Survivin shRNA interference technology was applied to liposomes transfection esophageal ECA109 cells,RT-PCR and Western blot methods was applied to evaluate genes expression of Survivin and CDK4. Flow cytometry was used to detect the proliferation of cell lines index changes after ECA109 esophageal cell transfection. RESULTS:After Survivin shRNA transfection of esophageal ECA109 cell,Survivin mRNA and protein expressions were obviously down-regulated. CDK4 mRNA expression was also down-regulated. Cell ratio was increased in G2 phase but decrease in S phase,with the cell cycle blocked. CONCLUSION:Survivin was effective on adjusting CDK4 gene expression at mRNA level. The role of silencing Survivin down-regulated expression of CDK4 mRNA,suggesting that Survivin could be a new gene regulatory factor in the tumor signal control network.