Induction of the 47 kDa platelet substrate of protein kinase C during differentiation of HL-60 cells

Abstract

Immunoblot analysis showed that the 47 kDa platelet substrate of protein kinase C (P47) was expressed at low levels in undifferentiated HL-60 leukaemia cells. Treatment of these cells with dimethyl sulphoxide, 1 alpha,25-dihydroxycholecalciferol or retinoic acid caused progressive increases in P47 content. Retinoic acid (1 microM) elicited the largest response, a 4-fold increase in P47 protein after 7 days that was accompanied by an increase in translatable P47 mRNA. The induction of P47 by retinoic acid preceded cessation of cell proliferation and development of the capacity to reduce Nitro Blue Tetrazolium, indicating that its expression is an early event in the myeloid differentiation of HL-60 cells.

FMLP-induced formation of F-actin in HL60 cells is dependent on PI3-K but not on intracellular Ca 2+ , PKC, ERK or p38 MAPK

“…HL60 cells were differentiated into neutrophil-like cells by growth medium containing 1.1% DMSO for 5 days [20,21]. The cell viability determined by trypan blue exclusion was more than 90 % after differentiation.…”

Molecular analysis of pleckstrin: The major protein kinase c substrate of platelets

“…It thus might be under different translational control than the longer transcripts, perhaps increasing expression of pleckstrin upon differentiation in a manner analogous to the HL-60 a-tubulin system [27]. However, pleckstrin appears to be regulated during HL-60 differentiation primarily at the level of transcription or mRNA stability since in all cases protein abundance, as detected by Western blotting, paralleled mRNA levels [4,10]. Increased expression of pleckstrin as HL-60 cells differentiate into mature white cells correlates with the tissue distribution of pleckstrin mRNA and protein [4] (Stewart et al, in preparation), suggesting pleckstrin may confer a function common to differentiated hemopoietic cells.…”

Metallothionein RNA levels in HL‐60 cells Effect of cadmium, differentiation, and protein kinase C activation

“…To induce differentiation, cultures were treated with 20 nM TPA or 1 nM TPA plus 400 nM A23187 for 2 days, or 100 nM calcitriol or 1 ,aM RA for 5 days, and monitored by standard procedures including morphology, cell cycle exit, adherence, and biochemical markers [16,20]. Stock solutions of TPA (1000 x in ethanol), RA (1000 x in ethanol), and calcitriol (1000 × in dimethylsulfoxide) were stored at -20°C.…”