Cyclin B1的启动子在HeLa细胞cyclin B1蛋白调控中的作用  

Role of promoter of cyclin B1 in regulation of cyclin B1 protein in HeLa cells

作　　者：王昱[1,2] 卢仲义 许峰 朱宇迪 朱宇熹 

机构地区：[1]第三军医大学西南医院儿科,重庆400038 [2]重庆庆医科大学附属儿童医院PICU,重庆400014 [3]重庆庆医科大学附属儿童医院药剂科,重庆400014 [4]重庆庆医科大学附属儿童医院肿瘤科,重庆400014 [5]Department of Radiation Oncology&Image-applied Therapy,Kyoto University Graduate School of Medicine,Japan

出　　处：《第三军医大学学报》2009年第24期2440-2443,共4页Journal of Third Military Medical University

摘　　要：目的通过检测细胞周期素B1(cyclinB1)的启动子在HeLa细胞的不同细胞周期的活性变化,探讨其对cy-clinB1蛋白的调控作用。方法采用PCR法获得HeLa细胞cyclinB1的启动子,通过基因重组插入pGL3promotervector从而获得质粒pGL3/cyclinB1promoter。采用羟基脲(hydroxyurea,HU)对HeLa细胞进行细胞周期G1、S、G2/M期的同步化,通过流式细胞术法确认。用短暂转染的方法,将重组的质粒转染至HeLa细胞。通过双萤光素酶活性检测分析cyclinB1的启动子在HeLa细胞周期G1、S、G2/M期的活性的变化。结果双荧光素酶活性检测cyclinB1的启动子活性在G1期最高,S期最低,G2期中等。结论在HeLa细胞中cyclinB1的启动子活性和cyclinB1蛋白的表达并不同步。Objective To investigate the activity of cyclin B1 promoter in different cell cycle of HeLa cells and to explore its regulation role on cyclin B1 protein. Methods The cyclin B1 promoter of HeLa cells was obtained by PCR amplification, and then the PCR product was inserted into the plasmid of pGL3 promoter vector by recombination to construct the plasmid of pGL3/cyclin B1 promoter. HeLa cells were synchronized in G1 , S, G2/M phase by hydroxyurea （HU） , which was then identified by flow cytometry. The plasmid of pGL3/cyclin B1 promoter was transiently transfected into HeLa cells. The dual luciferase assay was performed in the HeLa cells at G1 , S, or G2/M phase for the promoter activity. Results The activity of cyclin B1 promoter was highest in G1 phase, decreased in S phase, and was medium in G2/M phase. Conclusion The activity of cyclin B1 promoter is different from the expression of cyclin B1 protein in the cell cycles of HeLa cells.

关 键 词：细胞周期素B1 启动子 HELA细胞 荧光素酶 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学] R394-33[医药卫生—基础医学]