g(m3)-ganglioside and Leukemia--Promyelocytic--Acute

The patterns of glycosphingolipids (GSLs) were analyzed in an HL-60 cell variant, HL-205, which is susceptible to phorbol 12-myristate 13-acetate (PMA)-induced monocyte/macrophage differentiation, and in an HL-60 cell variant, HL-525, which is resistant to such differentiation. The amounts and types of the GSLs were similar in both the HL-205 and HL-525 cells and they resemble those of granulocytes. Treatment with 3 nM PMA caused the susceptible HL-205 cells (but not the resistant cells) to acquire a new GSL pattern which resembles that of monocytes. This new pattern was characterized by increases in the level of a neutral GSL, Gb3Cer, from trace levels to 0.05 mg/10(9) cells and of an acidic GSL, GM3 ganglioside, from 0.03 to 0.33 mg/10(9) cells. The increases in the level of this ganglioside were found to be due to an increase in CMP-N-acetylneuraminic acid:lactosylceramide sialyltransferase activity. These results indicate an association between PMA-induced terminal differentiation along the monocyte/macrophage cell lineage and PMA-evoked increases in specific GSLs, GM3 in particular, which is due to a rise in the activity of its synthetic enzyme.

Topics: beta-D-Galactoside alpha 2-6-Sialyltransferase; Carbohydrate Sequence; Cell Differentiation; Enzyme Induction; G(M3) Ganglioside; Glycosphingolipids; Humans; Leukemia, Promyelocytic, Acute; Molecular Sequence Data; Monocytes; Sialyltransferases; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured

Conditions were developed to prolong the ability of sphinganine, a potent inhibitor of protein kinase C, to block the phorbol ester-induced adherence of HL-60 cells beyond 24 h. The loss of inhibition after this time seen previously (A.H. Merrill, Jr., A.M. Sereni, V.L. Stevens, Y.A. Hannun, R.M. Bell, and J.M. Kinkade, Jr., J. Biol. Chem., 261: 12610-12615, 1986), which appeared to be due to metabolism of this long-chain base, was overcome by supplying sphinganine daily. After 4 days, phorbol myristate acetate-induced adherence was inhibited approximately 50% by sphinganine. Sphinganine significantly decreased the expression of nonspecific esterase induced by phorbol myristate acetate in the nonadherent cells, indicating that other aspects of maturation besides adherence were blocked. The effects of daily sphinganine treatments on the monocytic differentiation induced by 1 alpha-25-dihydroxyvitamin D3 or ganglioside GM3 were also investigated. The increases in nonspecific esterase expression, nitroblue tetrazolium reduction, and morphological maturation caused by either agent were unaffected by the long-chain base. In addition, the changes in several cell surface antigens caused by 1 alpha,25-dihydroxyvitamin D3 were unaltered by sphinganine. Although phorbol esters, 1 alpha,25-dihydroxyvitamin D3, and ganglioside GM3 all induce the maturation of HL-60 cells along the monocytic lineage, the finding that sphinganine only affected the differentiation initiated by phorbol esters, in which protein kinase C clearly is a major regulator, suggests that this enzyme does not play a major role in these other pathways of differentiation.

Topics: Antigens, Neoplasm; Antigens, Surface; Calcitriol; Cell Differentiation; Cell Division; Cell Line; Cell Survival; G(M3) Ganglioside; Gangliosides; Humans; Kinetics; Leukemia, Promyelocytic, Acute; Sphingosine; Tetradecanoylphorbol Acetate; Tumor Cells, Cultured