上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (3): 284-.

• 论著（基础研究） • 上一篇    下一篇

不同浓度Nocodazole对Hela细胞G2/M期同步化的调节作用

徐秋芳1, 余克花1, 易 婷1, 黎 帆1, 刘发娣2, 应 颖1, 邹伟文1, 何 平3, 黄孝天1   

  1. 1. 南昌大学 基础医学院微生物学教研室, |南昌 330006；2. 江西省儿童医院检验科, 南昌 330006；3. 上海交通大学 基础医学院病原生物学教研室, 上海 200025
  • 出版日期:2010-03-25 发布日期:2010-03-24
  • 通讯作者: 黄孝天, 电子信箱: xthuang@ncu.edu.cn。
  • 作者简介:徐秋芳（1985—）, 女, 硕士生；电子信箱: qiufang588@yahoo.com.cn。
  • 基金资助:

    国家自然科学基金（30660010）；江西省教育厅科技计划项目（2006323）；江西省科技厅2008年科技支撑项目

Regulation effects of Nocodazole with different concentrations on Hela cell synchronization in G2/M phase

XU Qiu-fang1, YU Ke-hua1, YI Ting1, LI Fan1, LIU Fa-di2, YING Ying1, ZOU Wei-wen1, HE Ping3, HUANG Xiao-tian1   

  1. 1. Department of Medical Microbiology, Basic Medical College, Nanchang University, Nanchang 330006, China；2. Department of Laboratory Science, Children's Hospital of Jiangxi Province, Nanchang 330006, China；3. Department of Medical Microbiology, Basic Medical College, Shanghai Jiaotong University, Shanghai 200025, China
  • Online:2010-03-25 Published:2010-03-24
  • Supported by:

    National Natural Science Foundation of China, 30660010；Science and Technology Foundation of Education Department of Jiangxi Province, 2006323；Science and Technology Foundation of Jiangxi Province, Year 2008

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摘要：

目的 观察不同浓度有丝分裂阻滞剂Nocodazole对Hela细胞株G2/M期同步化的调节作用及其对纺锤体结构的影响。方法 分别以3.0、1.0、0.3 μmol/L Nocodazole处理Hela细胞（Nocodazole 3.0、1.0、0.3 μmol/L处理组）18 h。于Nocodazole撤除后0（处理18 h）、3、6、9 h时间点收集细胞，流式细胞仪检测G2/M期细胞百分比；间接免疫荧光染色激光共聚焦显微镜观察细胞有丝分裂器纺锤体α-微管蛋白（α-tubulin）排列。以不添加Nocodazole处理的Hela细胞作为对照组。结果 Nocodazole处理18 h，Nocodazole 3.0、1.0、0.3 μmol/L处理组Hela细胞G2/M期细胞百分比分别为55.95%、51.09%和47.81%，均显著高于对照组的9.54%（P<0.05）。在Nocodazole撤除后3、6、9 h时间点，Nocodazole 3.0 μmol/L和1.0 μmol/L处理组G2/M期细胞百分比无明显变化；而Nocodazole 0.3 μmol/L处理组G2/M期细胞百分比下降明显（30.43%、12.91%、10.23%），撤除后6 h时间点的G2/M期细胞百分比与对照组比较差异已无统计学意义（P>0.05）。α-tubulin荧光染色激光共聚焦显微镜观察显示，Nocodazole撤除后Nocodazole 0.3 μmol/L处理组G2/M期细胞微管迅速再聚合形成两极纺锤体且纺锤丝结构清晰。结论 Nocodazole对Hela细胞G2/M期同步化具有调节作用；药物撤离后,0.3 μmol/L Nocodazole处理组细胞周期恢复迅速且对纺锤体结构影响最小。

关键词: Nocodazole, Hela细胞, 细胞周期, 纺锤体, G2/M期

Abstract:

Objective To investigate the regulation effects of Nocodazole with different concentrations on Hela cell synchronization in G2/M phase and spindle morphology. Methods Hela cells were treated with 3.0, 1.0 and 0.3 μmol/L Nocodazole, respectively for 18 h (Nocodazole 3.0, 1.0, 0.3 μmol/L group, respectively). Cells were harvested 0 h, 3 h, 6 h and 9 h after removal of Nocodazole, and the percentage of cells in G2/M phase was detected by flow cytometry. Besides, the arrangement of α-tubulin of cells was observed by confocal microscopy with indirect immunofluorescence staining. Hela cells without treatment with Nocodazole were served as controls. Results After treatment with Nocodazole for 18 h, the percentages of Hela cells in G2/M phase in Nocodazole 3.0, 1.0, 0.3 μmol/L group were 55.95%, 51.09% and 47.81%, respectively, and were significantly higher than that in control group (9.54%)（P<0.05）. There was no significant change in the percentages of Hela cells in G2/M phase in Nocodazole 3.0 and 1.0 μmol/L group 3, 6 and 9 h after removal of Nocodazole, while that in Nocodazole 0.3 μmol/L group decreased significantly （30.43%, 12.91% and 10.23%）, and there had been no significant difference in the percentage of Hela cells in G2/M phase with control group 6 h after removal of Nocodazole （P>0.05）. After removal of Nocodazole, microtubules of cells in G2/M phase in Nocodazole 0.3 μmol/L group reaggregated, and two-pole spindles were formed, with clear spindle fibers. Conclusion Nocodazole can regulate Hela cell synchronization in G2/M phase. After removal of Nocodazole, those treated with 0.3 μmol/L Nocodazole may recover better in cell cycle and spindle morphology.

Key words: Nocodazole, Hela cell, cell cycle, spindle, G2/M phase