分选后晚G_1期凋亡细胞Cyclin E表达分析  

Analysis of Cyclin E Expression by Western Blot after Cell Sorting

作　　者：王尊[1] 黄丹[1] 葛军娜[1] 肖田[1] 魏欣[1] 李小兰[1] 肖徽[1] 陶德定[1] 龚建平[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院肿瘤研究所,武汉430030

出　　处：《华中科技大学学报（医学版）》2009年第1期36-38,48,共4页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：卫生部临床学科重点项目(No.20012537);国家自然科学基金资助项目(No.30570908)

摘　　要：目的以G1期细胞凋亡为模型,检测细胞周期素Cyclin E是否与凋亡相关。方法收获紫外线照射后的人类急性淋巴细胞性白血病细胞,一部分采用流式细胞术将凋亡诱导后的细胞进行Cyclin E/DNA双参数分析和细胞周期特异性细胞凋亡API双参数分析;一部分利用流式细胞分选术将凋亡诱导后的细胞分为凋亡组和未凋亡组,Westernblot法检测并比较两组细胞Cyclin E蛋白表达情况。结果Cyclin E/DNA双参数分析观察到凋亡诱导后细胞周期阻滞过程中细胞Cyclin E表达水平明显升高。分选后Western blot检测观察到凋亡组Cyclin E表达明显低于未凋亡组。结论Cyclin E表达水平的升高可能对晚G期细胞的凋亡起到保护作用。Objective To find out whether Cyelin E is involved in apoptosis, based on the model of late G1 phase （GIL） apoptosis. Methods After irradiation by ultraviolet （UV）, human acute lymphatic leukemia MOLT-4 ceils were gathered. Later, one part of the ceils induced by UV was analyzed by Cyclin E/DNA multiparameter method and API multiparameter. The other part of the cells by cell sorting, was divided into 2 groups： the group of dead cells and the group of alive cells. Thereafter, the level of Cyclin E was compared between the two group by Western blot. Results The level of Cyclin E was increased after the inducement of the G1 apoptosis by Cyclin E/DNA multiparameter method. The level of Cyclin E in the group of dead cells was lower obviously than in the group of alive cells. Conclusion The elevation of Cyclin E expression level may protects the cells from GIL apoptosis.

关 键 词：晚G1期凋亡 细胞周期 细胞周期检测点 细胞分选术 细胞周期素 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]