The phosphatase inhibitors, orthovanadate and levamisole, inhibit induction of erythroid differentiation and abrogate the associated inhibition of glycolysis.
Alterations in protein tyrosine phosphate (PTP), lactate, and fructose 2,6-bisphosphate (F-2,6-P2) levels have been associated with induced MEL cell differentiation and commitment to terminal cell division (TCD). The possible relationships of perturbations in PTP metabolism and reduction in lactate formation during differentiation were investigated utilizing sodium orthovanadate, Na3VO4, primarily an inhibitor of PTP phosphatases, and levamisole, considered an alkaline phosphatase inhibitor. Both of these compounds were found to effectively inhibit the TCD-associated differentiation induced by DMSO, HMBA, and Na butyrate and to abrogate the differentiation-associated reduction in lactate accumulation due to these agents. However, they were found not to inhibit hemin-induced hemoglobin synthesis which is independent of TCD and does not alter lactate metabolism. Two brominated levamisole analogs, L-p-bromotetramisole and D-p-bromotetramisole, were also found to be inhibitors of TCD-associated differentiation and to be effective at even lower concentrations than levamisole. The changes in TCD-associated differentiation and lactate production exhibited the same concentration-dependence with respect to the inhibitors. These findings strengthened the theory that TCD-associated differentiation, decreased lactate production, and sensitivity to phosphatase inhibitors are all associated. Since the induction of MEL cell differentiation has been shown to be associated with, and is thought to be due to, the induction of PTP phosphatase activity and Na3VO4 is thought to inhibit the differentiation by inhibiting PTP phosphatase activity, the effect of levamisole on PTP levels was determined. Levamisole, like Na3VO4, was found to increase the tyrosine phosphate levels of proteins of similar molecular weights in intact cells in both the presence and absence of a differentiation inducer. Several phosphotyrosine-containing, similarly sized proteins were particularly affected by differentiation induction and by Na3VO4 and levamisole treatment. Changes in the levels of tyrosine phosphate-containing proteins of approximately 92-96, 60, and 38 kd were particularly noticeable. The induction of differentiation reduced PTP levels and inhibition of differentiation due to treatment with either Na3VO4 or levamisole increased their levels. These data suggest relationships between signal transduction pathways involved in differentiation and TCD, the regulation of lactate and F-2,6-P2 metabolism, and PTP levels.
Scher
B M
BM
Department of Microbiology, Mount Sinai Medical Center, Box 1194, 1 Gustave L. Levy Pl., New York, NY 10029, USA.
Fuksina
I
I
Hellinger
N
N
Waxman
S
S
Scher
W
W
D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D004920:Erythropoiesis / Q000187:drug effects / Q000502:physiology; D005635:Fructosediphosphates / Q000378:metabolism; D006019:Glycolysis / Q000187:drug effects / Q000502:physiology*; D007700:Kinetics; D007773:Lactates / Q000378:metabolism; D004915:Leukemia, Erythroblastic, Acute; D007978:Levamisole / Q000494:pharmacology*; D051379:Mice; D008954:Models, Biological; D017027:Protein Tyrosine Phosphatases / Q000037:antagonists & inhibitors*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013498:Suramin / Q000494:pharmacology; D013773:Tetramisole / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014638:Vanadates / Q000494:pharmacology*
D005635:Fructosediphosphates; D007773:Lactates; D007978:Levamisole; D014638:Vanadates; D013498:Suramin; C027652:fructose 2,6-diphosphate; D013773:Tetramisole; D017027:Protein Tyrosine Phosphatases; D004121:Dimethyl Sulfoxide