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中心体蛋白centrobin缺失对细胞周期的影响

发表时间:2011-09-15  浏览量:1230  下载量:398
全部作者: 王婵娟,陈秀婷,周星煜,田寒,戴婷,宋立兵,李隽
作者单位: 中山大学中山医学院;中山大学附属肿瘤医院
摘 要: 目的:研究centrobin在细胞周期中的生物学意义。方法:利用瞬时转染方法建立细胞模型,再通过免疫荧光、蛋白质印迹检测转染效率。利用MTT实验、溴脱氧尿苷(BrdUrd)掺入实验和免疫荧光检测细胞增殖,流式细胞技术检测细胞周期。结果:在乳腺癌细胞系MCF-7中,沉默centrobin基因表达阻止细胞从G1期进入S期,并且抑制细胞增殖。但是,在p53基因突变的乳腺癌细胞系MDA-MB231中,干扰centrobin的表达对细胞的增殖并无影响。进一步研究发现,在MCF-7细胞中敲除centrobin基因能够上调细胞周期调控因子p53的蛋白水平,导致细胞停滞在G1-S期。结论:centrobin在调控细胞周期进程的过程中扮演着重要角色,干扰centrobin表达导致的中心体复制障碍与细胞周期进程之间存在着密切的关系。
关 键 词: 分子生物学;细胞周期;centrobin;流式细胞技术;p53
Title: Effect of centrobin depletion on cell cycle
Author: WANG Chanjuan, CHEN Xiuting, ZHOU Xingyu, TIAN Han, DAI Ting, SONG Libing, LI Jun
Organization: Zhongshan School of Medicine, Sun Yat-sen University; Sun Yat-sen University Cancer Center
Abstract: Objective: To investigate the biological significance of centrobin in cell cycle. Methods: Si-RNA transfections were used to establish cell model. Immunofluorescence and western blotting were used to estimate the efficiency of the transfections. And, MTT assay, bromodeoxyuridine labeling and immunofluorescence were used to detect cell proliferation. Flow cytometry was used to analyze cell cycle. Results: Silencing centrobin expression prevented breast cancer cell line MCF-7 from G1 phase to S phase and inhibited MCF-7 proliferation, whereas another breast cancer cell line MDA-MB231, a p53 mutation cell line, was not affected. Furthermore, cell cycle regulator p53 was up-regulated while centrobin was knocked out in MCF-7 cells. Conclusion: Centrobin plays an important role in the regulation of cell cycle. Impaired centrosome duplication by interfering the expression of centrobin is closely related with the cell cycle progression.
Key words: molecular biology; cell cycle; centrobin; flow cytometry; p53
发表期数: 2011年9月第17期
引用格式: 王婵娟,陈秀婷,周星煜,等. 中心体蛋白centrobin缺失对细胞周期的影响[J]. 中国科技论文在线精品论文,2011,4(17):1549-1555.
 
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