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Departments of Biochemical Toxicology [L. Z., T. Y., Y. K.] and Biochemistry [K. N.], School of Pharmaceutical Sciences, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo 142, Japan
We have recently demonstrated that bufalin is a new potent inducer of the differentiation of human myeloid leukemia cells. The present work was carried out to examine further the effect of bufalin on the growth and characteristics of human leukemia-derived cell lines U937, ML1, and HL60. At concentrations of 5–10 nM, bufalin decreased the growth of ML1 cells preferentially at the G2 phase and U937 cells at the S and G2 phases of the cell cycle. Bufalin, under these conditions, induced the differentiation of U937, ML1, and HL60 cells to monocyte/macrophage-like cells by measuring the expression of various differentiation markers, as assessed by morphology and histochemistry, and ability to phagocytose latex particles, to reduce nitroblue tetrazolium, and to develop Fc receptors. U937 and ML1 cells started to differentiate at 4 and 6 h, respectively, after treatment with 10 nM bufalin and showed maximum differentiation 72 h later. At present, a mechanism for the bufalin-mediated induction of the differentiation of these human leukemia cells remains to be determined.
The combination of bufalin with all-trans retinoic acid, 1
,25-dihydroxyvitamin D3, 4'-demethylepipodophyllotoxin ethylidene-ß-D-glucoside (VP16), or human 
-interferon synergistically induced the differentiation of HL60 and U937 cells. A similar effect on ML1 cells was observed with the combination of bufalin with VP16 or human rTNF-.
These results suggest that bufalin in combination with VP16, all-trans retinoic acid, 1,25-dihydroxyvitamin D3, rTNF-, or -interferon may be very useful in the differentiation of human leukemia.
1 To whom requests for reprints should be addressed, at Tokyo Institute for Immunopharmacology, Inc., 3-41-8, Takada, Toshima-ku, Tokyo 171, Japan.
Received 1/ 2/92. Accepted 6/24/92.
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