Standard and quantitative analysis of cyclin E threshold by cyclin E/DNA multiparameter flow cytometry.
- Author:
Daxing, XIE
;
Yongdong, FENG
;
Jianhong, WU
;
Shuangyou, LIU
;
Xiaolan, LI
;
Deding, TAO
;
Jianping, GONG
- Publication Type:Journal Article
- MeSH:
Caffeine/pharmacology;
Cell Cycle/*physiology;
Cell Line, Tumor;
Cyclin E/*analysis;
Cycloheximide/pharmacology;
DNA, Neoplasm/*analysis;
Flow Cytometry/methods;
Jurkat Cells;
Leukemia, Lymphoid/pathology
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2005;25(3):282-4
- CountryChina
- Language:English
-
Abstract:
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.
