Effects of phorbol esters on an interleukin-3-dependent cell line.
FDC-P1 is an interleukin-3 (IL-3)-dependent cell line that ceases to proliferate in the absence of IL-3. We have isolated variant cell lines from FDC-P1 that grow in response to phorbol myristate acetate (PMA). These variant cell lines (FD/PMA) have maintained their PMA-dependency for over 1 year. Lymphokine gene expression, which would support growth, was not detected in FD/PMA lines. FD/PMA lines had a different cell surface phenotype than the parental line. Mac-1, Mac-2, and Mac-3 were readily detected on the cell surface of FD/PMA lines; however, these antigens were not detected on FDC-P1. Because protein kinase C (PKC) activation may mediate PMA effects, we examined this kinase. PKC activity quantitated by 32P-incorporation into histone was increased in FDC-P1 as compared with FD/PMA cultured in IL-3. Moreover, PKC activity was undetectable in FD/PMA lines cultured in PMA. Using Western blotting, immunoreactive PKC was readily detected in cytosolic and solubilized particulate fractions of FDC-P1 cells, not but in FD/PMA cell extracts. Comparisons between the parental and FD/PMA lines should provide insight into IL-3- and PMA-mediated signal transduction.
McCubrey
J A
JA
Department of Microbiology, East Carolina University School of Medicine, Greenville, NC.
Steelman
L S
LS
Sandlin
G
G
Riddle
R S
RS
Ways
D K
DK
D000818:Animals; D002460:Cell Line; D002462:Cell Membrane / Q000648:ultrastructure; D002469:Cell Separation; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology; D004357:Drug Synergism; D015870:Gene Expression / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D007377:Interleukin-3 / Q000502:physiology*; D008222:Lymphokines / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D010703:Phorbol Esters / Q000494:pharmacology*; D011493:Protein Kinase C / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013755:Tetradecanoylphorbol Acetate / Q000032:analysis / Q000494:pharmacology; D013936:Thymidine / Q000493:pharmacokinetics; D014316:Tritium
D003470:Culture Media; D007377:Interleukin-3; D008222:Lymphokines; D010703:Phorbol Esters; D012333:RNA, Messenger; D014316:Tritium; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate; D013936:Thymidine