A RAS oncogene imparts growth factor independence to myeloid cells that abnormally regulate protein kinase C: a nonautocrine transformation pathway.
The factor-dependent cell line FDC-P1 has been utilized as a model of interleukin 3 (IL-3)-dependent myeloid cell proliferation. However, it has been recently observed that active phorbol esters (e.g., phorbol 12-myristate 13-acetate) may entirely replace IL-3 to promote its proliferation. These observations reveal abnormal regulation of protein kinase C (pkC) (absence of downregulation or overexpression). This property allowed a test of the hypothesis that the T24 RAS (codon 12) oncogene acts by constitutive and persistent pkC activation, driving proliferation. FDC-P1 cells were transfected by electroporation with the T24 RAS-containing vector pAL 8, or with a control vector pSVX Zip Neo, and neomycin-resistant clones were selected. Multiple RAS-transfectant clones were categorized for their growth factor requirement and incorporation of the 6.6-kb human mutant H-RAS genome. IL-3-independent clones had incorporated multiple (more than two) copies of the entire 6.6-kb RAS genome. The incorporation of multiple 6.6-kb RAS genomes was correlated with high-level p21 RAS expression. No evidence for autostimulatory growth factor production by clones containing the RAS oncogene was observed. Thus, acquisition of growth factor independence in myeloid cells by abundant expression of a RAS oncogene is linked, in part, to abnormal regulation of pkC, which acts as a collaborating oncogene.
Department of Medicine, Indiana University School of Medicine, Indianapolis.
H S
HS
Boswell
T S
TS
Nahreini
G S
GS
Burgess
A
A
Srivastava
T G
TG
Gabig
L
L
Inhorn
E F
EF
Srour
M A
MA
Harrington
D002455:Cell Division; D002460:Cell Line; D002471:Cell Transformation, Neoplastic; D015536:Down-Regulation; D004351:Drug Resistance / Q000235:genetics; D004789:Enzyme Activation; D015870:Gene Expression; D011905:Genes, ras; D005822:Genetic Vectors; D006098:Granulocytes / Q000166:cytology* / Q000201:enzymology; D006801:Humans; D007377:Interleukin-3 / Q000494:pharmacology; D009154:Mutation; D009355:Neomycin; D011493:Protein Kinase C / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection
D007377:Interleukin-3; D011493:Protein Kinase C; D009355:Neomycin; D013755:Tetradecanoylphorbol Acetate