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 multiparamter 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.
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XIE Daxing, male, born in 1980, M. D., Ph. D.
This project was supported by grants from the National Natural Sciences Foundation of China (No. 39672065, 39730270, and 39725027), the China State Key Basic Research Program (No. G1998051212), and the Science Foundation of Ministry of Health, P. R. China (No. 20012537).
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Daxing, X., Yongdong, F., Jianhong, W. et al. Standard and quantitative analysis of cyclin E threshold by cyclin E/DNA multiparameter flow cytometry. Current Medical Science 25, 282–284 (2005). https://doi.org/10.1007/BF02828143
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DOI: https://doi.org/10.1007/BF02828143