基础医学与临床 ›› 2014, Vol. 34 ›› Issue (4): 470-474.

• 研究论文 • 上一篇    下一篇

NOK癌基因对人胚肾293T细胞周期G1/S期的影响及其作用机制

刘斌1,刘力2   

  1. 1. 中国医学科学院基础医学研究所
    2. 医科院基础医学研究所
  • 收稿日期:2013-12-23 修回日期:2014-01-06 出版日期:2014-04-05 发布日期:2014-03-31
  • 通讯作者: 刘力 E-mail:lliu@pumc.edu.cn
  • 基金资助:
    国家自然科学基金

The Influence and mechanism of the NOK oncogene on the G1/S Distribution of human embro kidney 293T Cells

1,   

  • Received:2013-12-23 Revised:2014-01-06 Online:2014-04-05 Published:2014-03-31

摘要: 目的 探讨NOK癌基因对人胚肾293T细胞周期G1/S期的影响,并研究NOK对G1/S期调控的作用机制。方法 用流式细胞仪对瞬时转染NOK及pcDNA3.0空载体的293T、293ET及HeLa细胞进行周期检测,并用免疫印迹法(Western blot)在293T细胞中,对其细胞周期G1/S期检验点调控蛋白cyclin D/E1及CDK2/4等蛋白水平进行检测。结果 瞬时转染NOK可使293T、293ET及HeLa细胞G1期较转染空载体pcDNA3.0的细胞比例减少,S期比例增多 (p<0.05)。转染NOK可使G1/S期检验点相关蛋白cyclin D、cyclin E1、p-CDK-2、p-CDK-4、p-Akt及p-Rb蛋白的表达量上升,使p21及p53蛋白表达量下降。结论 NOK可促进细胞快速通过细胞G1/S期检测点,其机制涉及对Akt及CDK2/4通路的影响。

关键词: NOK, 细胞周期, 周期素

Abstract: Objective To analyze the influence of the NOK oncogene on the G1/S distribution in 293T cells and to elucidate the regulated mechanisms responsible for NOK mediated G1/S phase control. Methods, After transiently transfecting NOK into 293T, 293ET and HeLa cells, we detected the cell cycle distributions by flow cytometric approach. Then, we detected the protein expressions of G1/S check point related genes such as cyclin D, cyclin E1,CDK2 and CDK4 by western blot analysis. Results In the 293T、293ET and HeLa cell which was transfered by NOK, the G1 phase proportions of cells were reduced, but the S phase proportions of cells were increased (p<0.05). Significant up-regulations in the expressions of cyclin D,cyclin E1, p-CDK-2, p-CDK-4, p-Akt and p-Rb were observed with the increased delivery of NOK into 293ET cells. Conclusion NOK can promote cell cycle progression by rapidly passing the G1/S check point, which may involve the activations of both Akt and CDK2/4 signaling pathways.

Key words: NOK, cell cycle, cyclin

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