Open AccessCharacterization of cyclin B1 expression in human cancer cell lines by a new three‐parameter BrdUrd/Cyclin B1/DNA analysis
Author(s) -
Faretta Mario,
Bergamaschi Daniele,
Taverna Stefano,
Ronzoni Simona,
Pantarotto Massimo,
Mascellani Enrico,
Cappella Paolo,
Ubezio Paolo,
Erba Eugenio
Publication year - 1998
Publication title -
cytometry
Language(s) - English
Resource type - Journals
eISSN - 1097-0320
pISSN - 0196-4763
DOI - 10.1002/(sici)1097-0320(19980101)31:1<53::aid-cyto7>3.0.co;2-k
Subject(s) - cyclin a , cyclin b1 , cyclin b , cell cycle , cyclin d , cyclin , biology , flow cytometry , cyclin e , cyclin a2 , microbiology and biotechnology , bromodeoxyuridine , cell , cell growth , biochemistry , cyclin dependent kinase 1
Flow cytometric cyclin expression/DNA content analysis, now commonly used, provides useful information on the mechanisms regulating cell cycle progression. However, this biparametric analysis does not make a clear‐cut distinction between G 1 and S‐early or between S‐late and G 2 M phase cells. This paper proposes a new three‐parameter flow cytometric method with which to determine cyclin B1 levels in single cells in different cell cycle phases by coupling bromodeoxyuridine (BrdUrd) immunodetection and DNA content. DNA denaturation by HCl did not alter the level of cyclin B1. Differences in cyclin B1 expression were observed in seven human cancer cell lines of different origin. The percentage of cyclin B1‐positive cells and the cyclin B1 content per cell indicated different patterns. In some cases cyclin B1 accumulation preceded the G 2 M checkpoint, at which its content usually started to rise. Using available easily reproducible techniques, this flow cytometric approach gives details of intracellular variability in cyclin expression. Cytometry 31:53–59, 1998. © 1998 Wiley‐Liss, Inc.