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Mitosis in Human Leukemic Leukocytes during Colcemid Inhibition and Recovery¹

Department of Biology, The University of Texas at Houston, M. D. Anderson Hospital and Tumor Institute and Graduate School of Biomedical Sciences, Houston, Texas 77025

Normal stimulated lymphocytes and peripheral leukocytes from five chronic and two acute myelogenous leukemia patients were treated with Colcemid, 0.02 g/ml, for 2 hr, arresting dividing cells in a C-metaphase configuration. Upon removal of Colcemid, cells progressed through metaphase, anaphase, and telophase during time intervals from 0 to 120 min. Normal and leukemic leukocytes in various stages of recovery were flat-embedded for analysis by electron microscopy. As in other mammalian cells, Colcemid inhibits the migration of centriole pairs to opposite poles and results in the formation of a unipolar spindle with chromosomes displaced radially about the centrioles. Upon removal of Colcemid, the centriole pairs moved apart and a normal bipolar spindle was formed. The recovery of normal stimulated lymphocytes proceeded rapidly up to 120 min. There was a gradual decrease in the percentages of C-metaphase cells with a subsequent increase in appearance of telophase cells.

The recovery of chronic myelogenous leukemia cells from Colcemid block proceeds at a rate much slower than normal lymphocytes. The percentages of telophase cells at 120 min was low and most of the cells at this point remained in C-metaphase. Mitotic abnormalities such as multipolar spindles, anaphase bridges, and lagging chromosomes were evident in some cells. Cells from three cultures of one patient with acute myelogenous leukemia failed to recover from Colcemid inhibition. Leukemic cells appeared to display a greater sensitivity to Colcemid. Specific defects were noted in the mitotic apparatus of leukemic leukocytes, including abnormal centriole migration and atypical fine structure.

¹ This work was supported in part by NIH Grants GM 15887, CA 05047, and CA 06939-05.

² To whom reprint requests should be addressed at Department of Human Biological Chemistry and Genetics, the University of Texas Medical Branch, Galveston, Texas 77550.

Received 10/15/71. Accepted 12/28/71.