Characterization of cyclin B1 expression in human cancer cell lines by a new three-parameter BrdUrd/cyclin B1/DNA analysis

Mario Faretta, Daniele Bergamaschi, Stefano Taverna, Simona Ronzoni, Massimo Pantarotto, Enrico Mascellani, Paolo Cappella, Paolo Ubezio, Eugenio Erba

Research output: Contribution to journalArticle

Abstract

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 G1 and S-early or between S-late and G2M 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 G2M check-point, 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.

Original languageEnglish
Pages (from-to)53-59
Number of pages7
JournalCytometry
Volume31
Issue number1
DOIs
Publication statusPublished - Jan 1 1998

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Keywords

  • Cyclin B1
  • G
  • Mitosis
  • Three-parameter flow cytometry

ASJC Scopus subject areas

  • Hematology
  • Cell Biology
  • Pathology and Forensic Medicine
  • Biophysics
  • Endocrinology

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