Kinetics of staurosporine-arrest of human diploid fibroblasts

Conference · Fri Jan 01 00:00:00 EST 1993 · Cytometry (Baltimore); (United States)

OSTI ID:6831614

Stevenson, A P; Bustos, L D; Dickson, J A; Habbersett, R C; Crissman, H A ^[1]

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1. Los Alamos National Lab., NM (United States)

The authors have previously shown that the general protein kinase inhibitor, staurosporine (Stsp), has a differential effect on the progression of normal and transformed cells. Low levels of Stsp (1.0 ng/ml) reversibly arrested normal diploid cells in early G1 phase, whereas concentrations of Stsp as high as 50-75 ng/ml had no effect on G1 progression in transformed cells. High concentrations of Stsp arrested normal cells in G1 and G2 phases but blocked transformed cells only in G2 phase of the cell cycle. To follow the accumulation of cells in G1 and/or G2 phases, asynchronous cultures of human diploid fibroblasts were treated with 0, 1, 5, 10, 20, 30 or 50 ng/ml staurosporine for 18 hours in the presence of bromodeoxyuridine. The kinetics of labeling with BrdU were determined with a two-laser flow cytometric technique that monitored emission of fluorescence from DNA-species stains that differed in the degree of BrdU-induced quenching of their fluorescence signals. Bivariate plots of Hoechst/mithramycin fluorescence signal ratio vs. Hoechst fluorescence identified the arrested cells as cohorts of cells in different positions across cell cycle at the time of Stsp addition.

• OSTI ID: 6831614
• Report Number(s): CONF-9303114-; CODEN: CYTODQ
• Journal Information: Cytometry (Baltimore); (United States), Vol. 6; Conference: 16. congress of the International Society for Analytical Cytology, Colorado Springs, CO (United States), 21-26 Mar 1993; ISSN 0196-4763
• Country of Publication: United States
• Language: English

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
CELL CYCLE
CONTROL
ENZYME INHIBITORS
QUANTITY RATIO
PHOSPHOTRANSFERASES
CYTOLOGICAL TECHNIQUES
FLUORESCENCE
LASERS
ONCOGENIC TRANSFORMATIONS
SORTING
CELL TRANSFORMATIONS
ENZYMES
LUMINESCENCE
ORGANIC COMPOUNDS
PHOSPHORUS-GROUP TRANSFERASES
PROTEINS
TRANSFERASES
550300* - Cytology
550200 - Biochemistry