26-26-26-27-27-27-hexafluoro-1-25-dihydroxyvitamin-d3 and Leukemia--Promyelocytic--Acute

The hexafluorinated vitamin D3 analog, 26,26,26,27,27,27-hexafluoro-1,25-dihydroxyvitamin D3[F6-1,25-(OH)2D3] is more potent than 1,25-dihydroxyvitamin D3 [1,25-(OH)2D3] regarding various physiological effects. When the biological potencies of vitamin D3 analogs were assessed 24 h after the addition by the induction of 24-hydroxylation activity in the human promyelocytic leukemia cell line, HL-60,F6-1,25-(OH)2D3 was 6 times as potent as 1,25-(OH)2D3 in a medium containing 5% fetal bovine serum. When the cells were cultured in a serum-free medium, F6-1,25-(OH)2D3 was only equipotent to 1,25-(OH)2D3. Considering a previous report demonstrating a weaker binding of F6-1,25-(OH)2D3 to serum vitamin D-binding protein (DBP) than 1,25-(OH)2D3, it seems that the resultant greater free fraction of F6-1,25-(OH)2D3 might account for its greater activity in a serum-containing medium. As assessed by the suppression of cell proliferation and the induction of cell differentiation along the monocyte/macrophage pathway which requires as long as 96 h for their assessment, the potency ratio of F6-1,25-(OH)2D3 to 1,25-(OH)2D3 increased as the levels of fetal bovine serum increased. It was of great interest that F6-1,25-(OH)2D3 was still significantly more potent than 1,25-(OH)2D3 even in a serum-free medium. Together with the data indicating the equipotency of F6-1,25-(OH)2D3 and 1,25-(OH)2D3 in the induction of 24-hydroxylation activity, it was suggested that decreased metabolic inactivation might contribute in part to the higher potency of F6-1,25-(OH)2D3 in the long-term effect.

Topics: Animals; Calcitriol; Cattle; Cell Differentiation; Cell Division; Culture Media; Fetal Blood; Humans; Hydroxylation; Leukemia, Promyelocytic, Acute; Time Factors; Tumor Cells, Cultured

Monocytic differentiation-inducing activity of 26,26,26,27,27,27-hexafluoro-1 alpha,25-dihydroxyvitamin D3 [26,27-F6-1 alpha,25-(OH)2D3] was re-evaluated in human promyelocytic leukemia (HL-60) cells in serum-supplemented or serum-free culture. The order of in vitro potency for reducing nitroblue tetrazolium (NBT) was 26,27-F6-1 alpha,25-(OH)2D3 greater than 1 alpha, 25-dihydroxyvitamin D3 [1 alpha,25-(OH)2D3] = 26,26,26,27,27,27-F6-1 alpha,23(S), 25-trihydroxyvitamin D3 [26,27-F6-1 alpha,23(S), 25-(OH)3D3] under serum-supplemented culture conditions, whereas the order was 1 alpha, 25-(OH)2D3 = 26,27-F6-1 alpha,25-(OH)2D3 greater than 26,27-F6-1 alpha,23(S), 25-(OH)3D3 under serum-free culture conditions. This rank order for differentiation-inducing activity under serum-free culture conditions correlated well with the binding affinity of these analogs for vitamin D3 receptor of HL-60 cells. The order of relative % binding affinity for the vitamin D-binding protein in fetal calf serum was 1 alpha,25-(OH)2D3 (100%) much greater than 26,27-F6-1 alpha,25-(OH)2D3 (5.1%) greater than 26,27-F6-1 alpha,23(S), 25-(OH)3D3 (less than 1%). These results suggest that serum vitamin D-binding proteins apparently modulate monocytic differentiation of HL-60 cells by 26,27-F6-1 alpha,25-(OH)2D3 under serum-supplemented culture conditions.

Topics: Binding, Competitive; Calcitriol; Cell Differentiation; Cell Line; Culture Media; Culture Media, Serum-Free; Humans; Kinetics; Leukemia, Promyelocytic, Acute; Receptors, Calcitriol; Receptors, Steroid; Vitamin D-Binding Protein