【摘要】 目的:探讨沉默核分裂周期蛋白80基因(nuclear division cycle 80,NDC80)对胆囊癌GBC-SD和NOZ细胞增殖、迁移和侵袭能力的影响。方法:应用实时荧光定量PCR和蛋白质印迹法分别检测NDC80 siRNA转入胆囊癌GBC-SD和NOZ细胞中后NDC80 mRNA和蛋白表达水平的变化;分别采用CCK-8法检和平板克隆形成实验检测沉默NDC80基因表达后胆囊癌细胞增殖和克隆形成能力的变化;应用Transwell小室实验检测沉默NDC80基因表达后胆囊癌细胞的迁移和侵袭能力的变化。结果:NDC80 siRNA(si-NDC80组)和阴性对照siRNA(si-NC组)成功转入胆囊癌GBC-SD和NOZ细胞;GBC-SD和NOZ细胞si-NDC80组中NDC80 mRNA及蛋白表达量均明显低于si-NC组(P值均<0.001)。GBC-SD和NOZ细胞在si-NDC80转染0～4 d时的增殖能力与si-NC阴性对照组相比,均明显下降(P值均<0.000 1);而且si-NDC80转染组的细胞迁移和侵袭能力与si-NC阴性对照组相比,亦明显降低(P值均<0.000 1)。结论:沉默NDC80基因表达可抑制人胆囊癌细胞的增殖、迁移和侵袭能力,提示NDC80基因在胆囊癌发生和发展中起到重要作用。更多还原

【Abstract】 Objective:To investigate the effects of silencing nuclear division cycle 80 (NDC80) gene expression on the proliferation,migration and invasion abilities of human gallbladder carcinoma GBC-SD and NOZ cells.Methods:Real-time fluorescent quantitative PCR and Western blotting were used to detect the expressions of NDC80 mRNA and protein in gallbladder carcinoma GBC-SD and NOZ cells after the transfection of NDC80 siRNA,respectively.After NDC80 gene silencing,the proliferation of GBC-SD and NOZ cells were detected by CCK-8 test and clone formation test,respectively.Transwell chamber assay was used to detect the migration and invasion abilities of gallbladder carcinoma cells after NDC80 gene silencing.Results:The NDC80 siRNA (si-NDC80 group) and the negative control (si-NC group) were successfully transfected into gallbladder carcinoma GBC-SD and NOZ cells,respectively.The expression levels of NDC80 mRNA and proteins in GBC-SD and NOZ cells transfected with si-NDC80 were significantly lower than those in si-NC group (all P < 0.001).Compared with the si-NC group,si-NDC80 transfection reduced the proliferation and clone formation abilities of GBC-SD and NOZ cells (all P < 0.001),and the migration and invasion abilities of GBC-SD and NOZ cells transfected with si-NDC80 were significantly decreased (all P < 0.001).Conclusion:Silencing the expression of NDC80 gene can inhibit the proliferation,migration and invasion of human gallbladder carcinoma cells,suggesting that NDC80 plays an important role in the occurrence and development of gallbladder carcinoma.更多还原