Xie Daxing, Feng Yongdong, Wu Jianhong, Liu Shuangyou, Li Xiaolan, Tao Deding, Gong Jianping
The Cancer Research Institute, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2005;25(3):282-4. doi: 10.1007/BF02828143.
The threshold of cyclin E expression at G1/S boundary is a characteristic feature of cell cycle progressing. In this study, we tried to develop a quantitative approach to analyze cyclin E threshold by multiparameter flow cytometry. The expression of cyclin E in exponentially growing MOLT-4 cells was detected under different photomultiplier tube (PMT) voltages by cyclin E/DNA multiparameter flow cytometry. Additionally, cyclin E was detected in cells which were treated with caffeine and cycloheximide (CHX) under the same PMT voltage. Moreover, the expression of cyclin E in MOLT-4 cells was compared with that in JURKAT cells. Cyclin E threshold was quantified by formula B2/AxC (A, B, C indicates the minimum, threshold, and maximum of cyclin E fluorescence intensity, respectively). Results showed that in MOLT-4 cells, cyclin E threshold calculated by formula B2/AxC was invariable under different PMT settings. It was decreased in cells treated with caffeine and remained changeless in cells treated with cycloheximide. Cyclin E threshold in JURKAT cells was much lower than that in MOLT-4 cells. It was suggested that Formula B2/AxC we firstly set up could be used to analyze cyclin E expression threshold quantitatively.
细胞周期蛋白E在G1/S边界的表达阈值是细胞周期进程的一个特征。在本研究中，我们试图开发一种定量方法，通过多参数流式细胞术分析细胞周期蛋白E阈值。通过细胞周期蛋白E/DNA多参数流式细胞术在不同光电倍增管（PMT）电压下检测指数生长的MOLT-4细胞中细胞周期蛋白E的表达。此外，在相同PMT电压下，检测用咖啡因和放线菌酮（CHX）处理的细胞中的细胞周期蛋白E。此外，比较了MOLT-4细胞和JURKAT细胞中细胞周期蛋白E的表达。细胞周期蛋白E阈值通过公式B2/AxC进行量化（A、B、C分别表示细胞周期蛋白E荧光强度的最小值、阈值和最大值）。结果表明，在MOLT-4细胞中，通过公式B2/AxC计算的细胞周期蛋白E阈值在不同PMT设置下不变。在用咖啡因处理的细胞中它降低，在用放线菌酮处理的细胞中保持不变。JURKAT细胞中的细胞周期蛋白E阈值远低于MOLT-4细胞中的。提示我们首次建立的公式B2/AxC可用于定量分析细胞周期蛋白E表达阈值。
