IL-3 and ribavirin induce high level expression of megakaryocytic markers and messages during long-term treatment of a megakaryocytic leukemia cell line.
Megakaryocyte differentiation is a lengthy process with cells moving through a continuum delineated by the sequential expression of specific gene products. The limited number of primary cells available from marrow for analysis has brought attention to some leukemic cell lines which show enhanced megakaryocyte marker expression following incubation with inducing agents, the most common of which is phorbol myristate acetate (PMA). We developed an alternative induction protocol for the megakaryocytic leukemic cell line CMK, which involved incubation of the cells with IL-3 and the nucleoside analog, ribavirin, for 1-2 weeks. This treatment was neither toxic nor cytostatic and yielded increased levels of the surface glycoproteins GPIIb/IIIA and GPIb-IX. Levels of some megakaryocytic messages (GPIIIa, GPIX) showed a marked rise by 12 days of incubation in the inducer combination. This was due to a synergistic interaction between IL-3 and ribavirin which influenced both transcriptional and posttranscriptional events. Light and electron microscopy demonstrated the presence of large polyploid cells, with morphological features similar to those of megakaryocytes, in the induced cultures. Analysis of the heterogeneity of response in the cell population to the induction regimen after several days of treatment suggested that cells which failed to display surface markers had been stimulated by the inducers but did not have sufficient time to complete expression of that marker. The results were consistent with the view that the cells in the starting population were distributed along a temporal expression pathway, and those which were first to express the earliest marker would also lead in the expression of a later marker. The order of expression was the same as that during normal megakaryocyte development.
Otsuka Pharmaceuticals, Rockville, Maryland 20850.
A
A
Majumdar
S
S
Kerby
P E
PE
Stenberg
B
B
Mullikin
J H
JH
Beckstead
D A
DA
Cooney
M M
MM
Seidman
D000954:Antigens, Surface / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D002455:Cell Division / Q000187:drug effects; D004273:DNA, Neoplasm / Q000032:analysis / Q000235:genetics; D004357:Drug Synergism; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology*; D008533:Megakaryocytes / Q000737:chemistry / Q000378:metabolism / Q000473:pathology*; D008970:Molecular Weight; D010980:Platelet Membrane Glycoproteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D012254:Ribavirin / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013920:Thrombocythemia, Essential / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D013997:Time Factors; D014158:Transcription, Genetic; D014407:Tumor Cells, Cultured
D000954:Antigens, Surface; D004273:DNA, Neoplasm; D007377:Interleukin-3; D010980:Platelet Membrane Glycoproteins; D012333:RNA, Messenger; D012254:Ribavirin; D013755:Tetradecanoylphorbol Acetate