A rise and fall in 1,2-diacylglycerol content signal hexamethylene bisacetamide-induced erythropoiesis.
Previous studies have suggested a role for protein kinase C (PKC) during induction of murine erythroleukemia cell (MELC) differentiation by hexamethylene bisacetamide (HMBA) (Melloni, E., Pontremoli, S., Viotti, P. L., Patrone, M., Marks, P. A., and Rifkind, R. A. (1989) J. Biol. Chem. 264, 18414-18418). The present studies assess the effect of HMBA on the content of 1,2-diacylglycerol (DG), the physiologic activator of PKC, in MELC variants. Exposure of parental Sc9 cells to HMBA induced a rapid rise and fall in DG content. The DG level increased within seconds from 225 pmol.10(6) cells-1 to a maximum of 305 pmol.10(6) cells-1 at 5 min. Thereafter, DG content fell reaching control levels at 30 min and 46% of control at 4 h. Similar DG elevations were detected in HMBA-resistant, phorbol ester-resistant, and vincristine-resistant MELC lines. Early DG elevation was followed by the characteristic rapid fall in both the phorbol ester-resistant and vincristine-resistant lines, both of which differentiate rapidly in response to HMBA. In contrast, in an HMBA-resistant MELC the DG level failed to fall for at least 10 h. Selection of HMBA-resistant cells for vincristine resistance restores both HMBA sensitivity and the rapid fall in DG content. Addition of a synthetic DG, 1-oleyl-2-acetyl glycerol (OAG), along with HMBA and every 2 h for the next 48 h blocked differentiation, as measured by accumulation of benzidine-reactive cells or by the commitment assay in methyl-cellulose. However, if addition of OAG was delayed for just a few minutes, until endogenous DG levels began to fall, differentiation was no longer inhibited. Rapid elevation of DG content is the earliest reported event during HMBA action and a subsequent fall in the DG content appears to be a critical step in the process of commitment to terminal differentiation.
Michaeli
J
J
DeWitt Wallace Research Laboratory, Memorial Sloan-Kettering Cancer Center, New York, New York.
Busquets
X
X
Orlow
I
I
Younes
A
A
Colomer
D
D
Marks
P A
PA
Rifkind
R A
RA
Kolesnick
R N
RN
D000081:Acetamides / Q000494:pharmacology*; D000818:Animals; D004075:Diglycerides / Q000378:metabolism* / Q000494:pharmacology; D004351:Drug Resistance; D004920:Erythropoiesis / Q000187:drug effects*; D014644:Genetic Variation; D007700:Kinetics; D004915:Leukemia, Erythroblastic, Acute; D051379:Mice; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D014750:Vincristine / Q000494:pharmacology
D000081:Acetamides; D004075:Diglycerides; D014750:Vincristine; C038237:1-oleoyl-2-acetylglycerol; C014026:hexamethylene bisacetamide; D013755:Tetradecanoylphorbol Acetate