Induction of leukotriene C4 synthase after the differentiation of rat basophilic leukemia cells with retinoic acid and a low dose of actinomycin D and its suppression with methylprednisolone

Abstract

Leukotriene C4 synthase (LTC4 S) is a pivotal enzyme for generation of cysteinyl-leukotrienes (cysLTs). LTC4 S activity in rat basophilic leukemia-1 (RBL-1) cells increased after culture in the presence of retinoic acid (RA) analogues, which was inhibited by cycloheximide or actinomycin D (ACD). Unexpectedly, the co-addition of a low dose of ACD with RA further potentiated the upregulation of the LTC4 S activity. Daunorubicin and mitomycin C also had a similar effect. When stimulated with calcium ionophore A23187, control cells did not produce cysLTs, but RA-treated cells generated cysLTs and the co-addition of ACD further increased. While LTC4 S mRNA and protein increased in the cells treated with RA, the co- addition of ACD further potentiated both in proportion to the LTC4 S activity. The effect of ACD was considered to enhance the transcription rate of LTC4 S gene, but not the mRNA-stability. The addition of methylprednisolone (MP) inhibited generation of cysLTs from the cells with A23187-stimulation and also did LTC4 S activity, but did not inhibit 5-lipoxygenase (5-LOX). The suppression of LTC4 S with MP showed a dependent manner on the time-point and duration of MP-treatment after RA-addition which was correlated with reduction in LTC4 S mRNA and protein. The cells cultured with RA plus ACD contained more histamine, chymase activity, and granules in the cytoplasm than the control cells, suggesting differentiation to mature mast cells. In consideration of RA-differentiation therapy, it may be of pathophysiological relevance that the antineoplastic agents potentiate RA-induced, steroid-sensitive, induction of LTC4 S in RBL-1 cells. © 2003 Wiley-Liss, Inc.