Standard and Quantitative Analysis of Cyclin E Threshold by Cyclin E/DNA Multiparameter Flow Cytometry
- Author:
Daxing XIE
1
;
Yongdong FENG
;
Jianhong WU
;
Shuangyou LIU
;
Xiaolan LI
;
Deding TAO
;
Jianping GONG
Author Information
1. Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology
- Keywords:
cyclin E;
threshold;
quantitative analysis
- From:
Journal of Huazhong University of Science and Technology (Medical Sciences)
2005;25(3):282-284
- CountryChina
- Language:Chinese
-
Abstract:
Summary: 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/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 B2/A×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. It was suggested that Formula B2/A×C we firstly set up could be used to analyze cyclin E expression threshold quantitatively.
