Standard and Quantitative Analysis of Cyclin E Threshold by Cyclin E／DNA Multiparameter Flow Cytometry

• Published in: Current medical science 2005, Vol.25 (3), p.282-284
• Main Author: 谢大兴 冯永东 吴剑宏 刘双又 李小兰 陶德定 龚建平
• Format: Article
• Language: English
• Subjects: Caffeine - pharmacology; Cell Cycle - physiology; Cell Line, Tumor; Cyclin E - analysis; Cycloheximide - pharmacology; DNA; DNA, Neoplasm - analysis; Flow Cytometry - methods; Humans; Jurkat Cells; Leukemia, Lymphoid - pathology; MOLT-4; 标准定量分析; 环己酰亚胺; 环氧合酶E; 血细胞计数
• ISSN: 1672-0733; 2096-5230; 1993-1352; 2523-899X
• 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 photomuhiplier tube (PMT) voltages by cyclin E/DNA muhiparameter 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 B^2/A>(C (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 B^2/AX C 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, h was suggested that Formula B^2/A>(C we firstly set up could be used to analyze cyclin E expression threshold quantitatively.