Cellular ras oncogene expression and cell cycle measured by flow cytometry
in hematopoietic cell lines
M Andreeff, DE Slater, J Bressler and ME Furth
Human hematopoietic malignancies provide an excellent model for the study
of the activity of cellular oncogenes in a context of known defects in cell
proliferation and differentiation. A flow cytometric immunofluorescence
assay was developed to quantitate the expression of the cellular ras
oncogenes in relation to the cell cycle in individual leukemic cells.
Specific binding of a monoclonal antibody to the 21-kd protein (p21ras)
encoded by the Ha-ras, Ki-ras, and N-ras genes was measured by flow
cytometry and confirmed by fluorescence microscopy. P21ras was detected in
416B, a murine hematopoietic precursor cell characterized by a high level
of Ki-ras expression, and in the human leukemic cell lines P-12 and KG-1.
The presence of p21ras in the cell lines was also shown by
immunoprecipitation. Cellular DNA content was determined simultaneously to
define cell cycle phases. There was an equal distribution of p21ras in G1,
S, and G2M, with considerable heterogeneity of ras gene expression in the
G1 compartment. The assay allows oncogene expression to be studied in
populations of intact single cells in which cell heterogeneity is
maintained, requires very few cells per sample, and directly correlates
oncogene expression to cell kinetic data.
Volume 67,
Issue 3,
pp. 676-681,
03/01/1986
Copyright © 1986 by The American Society of Hematology
