靶向沉默EphA8基因对人结肠腺癌细胞凋亡及细胞周期的影响  

The effect of targeted silent gene EphA8 to cell apoptosis and cell cycle of colonic adenocarcinoma

作　　者：朱剑军[1] 段劼[2] 林雪梅[3] 王晓玲[3] 朱晓华[1] 谢兴国[1] 邓世山[1] 

机构地区：[1]川北医学院人体解剖学教研室 [2]川北医学院附属医院神经外科 [3]川北医学院附属医院病理科,四川南充637000

出　　处：《川北医学院学报》2014年第2期141-146,共6页Journal of North Sichuan Medical College

基　　金：四川省教育厅基金项目(13ZB0242);川北医学院科研发展计划项目(CBY12-A-QN01)

摘　　要：目的:通过转染siRNA片段,研究EphA8基因沉默后对结肠腺癌Ls174T细胞凋亡和细胞周期的影响。方法:设计合成三对靶向沉默EphA8基因表达的siRNA片段,通过Lipofectamine2000介导转染人结肠腺癌细胞Ls174T。采用实时荧光定量PCR技术和Western Blot检测EphA8基因沉默效率,流式细胞术检测细胞凋亡和细胞周期。结果:三对siRNA片段均能沉默结肠腺癌Ls174T细胞EphA8 mRNA的表达,siRNA3抑制效果最明显,抑制率为85.71%。流式细胞术检测细胞凋亡结果显示:转染48 h后,Ls174T细胞实验组细胞凋亡率为33.29%;流式细胞术检测细胞周期结果显示:转染60 h后,结肠腺癌Ls174T细胞实验组S期细胞和G2期细胞比例显著降低。结论:本实验成功筛选出特异性靶向沉默EphA8基因的siRNA片段,并初步鉴定了EphA8基因的功能。Objective：To study the effects of cell apoptosis and cell cycle after silencing the EphA8 gene through transferring the siR NA fragments into the colonic adenocarcinima Ls174T cells. Methods：Three pairs of targeted silent gene EphA8 expressed by siRNA fragments were designed and transported to the colonic adenocarcinoma Ls174T cells by transferring Lipofectamine2000. The real-time PCR and Western Blot were used to detect the silent efficiency,and the cell apoptosis and cell cycle were measured by flow eytometry. Results ： All of the three pairs of siRNA can silent the gene expression of the EphA8 mRNA,and the suppression effect of siRNA3 was the most effective one. The inhibition rate was 85.71%. The result of cell apoptosis examined by flow cytometry showed that the rate of the Ls174T ceils was 33.29% after 48 hours of transferring,and the S stage and G2 stage cells were significantly decreased after trans- ferring the siRNA3 into the Ls174T cells through flow cytometry 60 hours later. Conclusion：The most effective siRNA fragments were chosen successfully for silencing the homologous EphA8 mRNA,and preliminarily identified its gene function.

关 键 词：EphA8 结肠腺癌 细胞凋亡 

分 类 号：R735.35[医药卫生—肿瘤]