Significance of proteolytic activity in 1,25-(OH)2D3-induced differentiation of HL-60 cells.

Two types of HL-60 cells, 1,25-(OH)2D3-responsive ATCC HL-60 cells and 1,25-(OH)2D3-resistant LG HL-60 cells were used. Despite the presence of enough amounts of normal 1,25-(OH)2D3 receptors, only 22% of LG cells matured after a 4-day treatment with 10(-7) M 1,25-(OH)2D3, while 80% of ATCC cells differentiated. However, 1,25-(OH)2D3 inhibited the proliferation of LG cells to the same degree as that of ATCC cells. 1,25-(OH)2D3 also induced (1) the ability to metabolize 1,25-(OH)2D3 to 1,24,25-(OH)3D3, and (2) up-regulation of the 1,25-(OH)2D3 receptor in LG as well as ATCC cells. Furthermore, the proportion of mature LG cells was 78% after treatment with 10(-7) M 1,25-(OH)2D3 for the first 48 h and 10(-7) M dbcAMP for the second 48 h, which was greater than that when treated only with 10(-7) M dbcAMP for the second 48 h (24.2%). These results indicate that 1,25-(OH)2D3 receptor complexes function normally in LG cells at commitment step in cell differentiation. ATCC cells had a serine proteinase to destroy specific 1,25-(OH)2D3-binding activity of the unoccupied receptor and digest 53-kD receptor to a small fragment with a MW of 16.3 kD, while not affecting the level of the specific binding of the occupied receptor. Other cells, such as murine leukemia cells, M1, and human chronic myeloid leukemia cells, the differentiation of which is induced by 1,25-(OH)2D3, seemed to have the same type of proteinase, suggesting the physiological significance of this proteinase in 1,25-(OH)2D3-induced cell differentiation.