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Induction of Differentiation of HL-60 Cells by Dimethyl Sulfoxide: Evidence for a Stochastic Model Not Linked to the Cell Division Cycle¹

Laboratory of Tumor Cell Biology, National Cancer Institute, NIH, Bethesda, Maryland 20205 [C. T., F. W. R.], and Cattedra di Ematologia and Cattedra di Patologia Sp. Medica, Universitá di Torino, Torino, Italy [D. F., E. G., G. L. P.]

Induction of differentiation of the human promyelocytic leukemia cell line HL-60 by dimethyl sulfoxide (Me₂SO) was analyzed to determine the relationship between exposure time of the inducer and cell cycle. A minimum incubation time of 12 hr with Me₂SO was required in order to induce differentiation in a small but significant proportion of cells. These expressed differentiation markers (morphology, phagocytosis, and nitroblue tetrazolium reduction) up to 12 hr after Me₂SO was removed from the medium. For periods beyond 12 hr and as long as 120 hr of contact of HL-60 cells with the inducing agent, a linear rise in the percentage of differentiated cells was observed. The sensitivity to Me₂SO of HL-60 cells synchronized by double thymidine block was examined and was found to be similar to that of nonsynchronized cells. The effect of Me₂SO was not altered when incubated with cells at different phases of the cell cycle. Finally, even nonproliferating cells were sensitive to the inducing effect of Me₂SO. The data are consistent with a stochastic model of induction to differentiation without having any linkage to the cell cycle.

¹ Supported in part by Regione Piemonte and S. Giovanni Hospital, Torino, Italy, and by CNR Grant 780283596, Progetto Finalizzato per la Crescita Neoplastica.

² Present address: Cattedra di Ematologia, Universitá di Torino, Torino, Italy.

³ To whom requests for reprints should be addressed.

Received 7/ 8/81. Accepted 10/27/81.

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