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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 the proliferation, cell cycle, Aurora-A, γ-tubulin and P53 protein expression in CasKi cells and its mechanism. Methods Caski cells were treated with different concentrations of nocturne. Cell proliferation was measured by MTT assay. Cell cycle was analyzed by flow cytometry. The changes of centrosomes were detected by indirect immunofluorescence. The expression of Aurora-A mRNA was detected by RT-PCR. Method to detect the expression of γ-tubulin, Aurora-A and P53 protein. RESULTS: Nordy significantly inhibited the proliferation of CasKi cells and blocked the cell cycle at G0 / G1 phase, and showed dose-dependent and time-dependent manner. The fluorescence signal of γ-tubulin in the center of cells was significantly reduced, and the expression of Aurora-A mRNA γ-tubulin and Aurora-A protein expression decreased significantly, while the P53 protein expression was significantly increased, and in a dose-dependent manner. CONCLUSION: Nordy may up-regulate P53 by down-regulating Aurora-A and exert its effect on inhibiting the proliferation of CasKi cells, arresting the cell cycle, preventing centrosome amplification and limiting the disorderly proliferation of cells.