不同浓度Nocodazole对Hela细胞G_2/M期同步化的调节作用  

Regulation effects of Nocodazole with different concentrations on Hela cell synchronization in G_2/M phase

作　　者：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] 

机构地区：[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　　处：《上海交通大学学报（医学版）》2010年第3期284-287,共4页Journal of Shanghai Jiao tong University:Medical Science

基　　金：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~~

摘　　要：目的观察不同浓度有丝分裂阻滞剂Nocodazole对Hela细胞株G_2/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、0.3μmol/L Nocodazole处理Hela细胞(Nocodazole 3.0、1.0、0.3μmol/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G_2/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、0.3μmol/L处理组Hela细胞G_2/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 3.0μmol/L和1.0μmol/L处理组G_2/M期细胞百分比无明显变化;而Nocodazole 0.3μmol/L处理组G_2/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G_2/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 0.3μmol/L处理组G_2/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G_2/M期同步化具有调节作用;药物撤离后,0.3μmol/L Nocodazole处理组细胞周期恢复迅速且对纺锤体结构影响最小。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 p,moL/L Noeodazole, respectively for 18 h （ Nocodazole 3.0, 1.0, 0.3 umol/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 eytometry. Besides, the arrangement of α-tubulin of cells was observed by confoeal microscopy with indirect immunofluorescence staining. Hela cells without treatment with Noeodazole were served as controls. Results After treatment with Noeodazole for 18 h, the percentages of Hela cells in G2/M phase in Nocodazole 3.0, 1.0, 0.3 umol/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 umol/L group 3, 6 and 9 h after removal of Nocodazole, while that in Nocodazole 0.3 umol/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 umol/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 umol/L Nocodazole may recover better in cell cycle and spindle morphology.

关 键 词：NOCODAZOLE HELA细胞 细胞周期 纺锤体 G2/M期 

分 类 号：R73-3[医药卫生—肿瘤]