诺帝对CasKi细胞增殖及中心体Aurora-A和γ-tubulin基因表达的影响  

Effect of Nordy on Proliferation of CasKi Cells and Expressions of Centrosome Aurora-A and γ-Tubulin Genes

作　　者：巫静娴[1] 赵涌[1] 卞修武[1] 

机构地区：[1]重庆医科大学分子医学与肿瘤研究中心病理教研室,重庆400016

出　　处：《中国生物制品学杂志》2011年第7期828-831,838,共5页Chinese Journal of Biologicals

摘　　要：目的探讨诺帝对CasKi细胞增殖、细胞周期及中心体Aurora-A、γ-tubulin和P53蛋白表达的影响及其作用机制。方法用不同浓度的诺帝处理CasKi细胞,MTT法检测细胞增殖活力,流式细胞术分析细胞周期,间接免疫荧光检测中心体的变化,RT-PCR检测细胞Aurora-A基因mRNA的表达,Western blot法检测细胞γ-tubulin、Aurora-A和P53蛋白的表达。结果诺帝能明显抑制CasKi细胞增殖,阻滞细胞周期于G0/G1期,且呈明显的剂量和时间依赖性;细胞中心体内γ-tubulin荧光信号明显减弱,Aurora-A基因mRNA的表达量及γ-tubulin和Aurora-A蛋白的表达量明显下降,而P53蛋白的表达量明显升高,且呈剂量依赖性。结论诺帝可能通过下调Aurora-A,上调P53,发挥其抑制CasKi细胞增殖、阻滞细胞周期,并阻止中心体扩增,限制细胞无序增殖的作用。Objective To investigate the effect of Nordy on proliferation and cell cycle of CasKi cells and expressions of centrosome Aurora-A and γ-tubulin genes as well as the relevant mechanism.Methods CasKi cells were treated with Nordy at various concentrations,and determined for proliferative activity by MTT method,for cell cycle by flow cytometry（FCM）,for change of centrosomes by IFA,for expressions of Aurora-A mRNA by RT-PCR,and for expressions of γ-tubulin,Aurora-A and P53 proteins by Western blot.Results Nordy significantly inhibited the proliferation of CasKi cells and arrested the cell cycle at G0/G1 stage,both in dose-and time-dependent modes.The fluorescence signal of γ-tubulin in centrosome was weakened significantly.The expression levels of Aurora-A mRNA as well as γ-tubulin and Aurora-A proteins decreased significantly,while that of P53 protein increased significantly,each in a dose-dependent mode.Conclusion Nordy might inhibit the proliferation of CasKi cells,arrest cell cycle,prevent the amplification of centrosomes and restrict the disordered proliferation of cells by down-regulating Aurora-A expression and up-regulating P53 expression.

关 键 词：诺帝 宫颈肿瘤 细胞增殖 中心体 AURORA-A Γ-TUBULIN 

分 类 号：R737.33[医药卫生—肿瘤] R979.1[医药卫生—临床医学]