›› 2010, Vol. 30 ›› Issue (3): 284-.

• Original article (Basic research) • Previous Articles     Next Articles

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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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