Control of lysozyme gene expression in differentiating HL-60 cells

We have investigated the control of lysozyme gene expression in HL-60 cells induced to differentiate into macrophage-like cells with phorbol myristate acetate (PMA). Differentiation, as evidenced by cellular adherence, and morphological changes corresponded temporally to an increase in nonspecific esterase activity. The lysozyme concentration in the medium of uninduced HL-60 cells was 10 Μg/10 7 cells. increasing to a maximum of 46 Μg/10 7 cells after 48 h incubation with PMA (16 nm). At 72 h the lysozyme concentration decreased to 16 Μg/10 7 cells. Intracellular lysozyme activity remained constant throughout differentiation. If HL-60 cells were exposed to PMA for 24 h, washed, then maintained in normal medium, they differentiated normally, confirming their irreversible commitment to differentiate. The increase in lysozyme secretion by these cells, however, is markedly blunted suggesting that continued PMA treatment of differentiated cells is required for their secretion of lysozyme. There is no change in the rate of extracellular degradation of lysozyme during differentiation. The level of lysozyme mRNA does not correlate directly with the amount of lysozyme secreted into the medium. Hybridization of uninduced HL-60 cell RNA with a chicken lysozyme cDNA probe demonstrates moderate hybridization. There is a modest (five-fold) increase in lysozyme mRNA between 0 and 36 h of exposure to PMA, corresponding to the burst of lysozyme secretion by these cells. The lysozyme mRNA decreases to a level which is lower than the original baseline by 72 h, when the cells are still secreting substantial amounts of lysozyme. These data suggest that both transcriptional and post-transcriptional controls are operative in the control of lysozyme gene expression during the differentiation of HL-60 cells. They also imply that lysozyme secretion is not a necessary component in the macrophage-monocyte differentiation of these cells.Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/71582/1/j.1365-2141.1985.tb07380.x.pd

All-trans retinoic acid-induced downregulation of annexin II expression in myeloid leukaemia cell lines is not confined to acute promyelocytic leukaemia

Most acute promyelocytic leukaemia (APL) patients suffer from disordered haemostasis. APL can be treated successfully in most instances by all-trans retinoic acid (ATRA) therapy, which induces endpoint maturation of the leukaemic promyelocytes with the characteristic t(15;17). Annexin II (AnII), a profibrinolytic protein, has been implicated in the bleeding manifestation seen in APL. Our group has shown previously that high levels of AnII are expressed on other acute myeloid leukaemia subtypes that are sometimes associated with disordered haemostasis, albeit less frequently than APL. This study examined the effects of ATRA on AnII expression and cell differentiation, on myeloid leukaemia cell lines to determine whether a regulatory influence on AnII may contribute to the return of haemostatic stability in APL following treatment. The results confirmed that AnII expression in the APL cell line (NB4) was significantly downregulated in response to ATRA (P < 0·01), with associated morphological and immunophenotypical evidence of myeloid differentiation. ATRA also downregulated AnII expression on other myeloid cell lines, albeit to a lesser extent than observed on NB4 cells. The results provide evidence that ATRA may resolve the hyperfibrinolysis in APL by downregulation of AnII expression