A mutant RAS gene acts through protein kinase C to augment interleukin-3 dependent proliferation in a fastidious immortal myeloid cell line

H. S. Boswell, M. A. Harrington, G. S. Burgess, T. L. Nahreini, H. G. Derigs, T. D. Hodges, D. English, C. D. Crean, T. G. Gabig

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Abstract

The functional role of a mutant RAS gene in immortal myeloid cell proliferation was examined in a fastidious interleukin-3 (IL-3) dependent cell line (NFS/N1.H7) formed by forced proliferation in IL-3 of marrow cells of the NFS/N mouse. The NFS/N1.H7 cell line was strictly dependent upon IL-3 for growth, and the cell line could be activated by phorbol esters (PMA) to augment IL-3 dependent proliferation, but when KC was downregulated, diminished IL-3 proliferative responses resulted. Transfection (electroporation) of the T24 RAS-containing vector pAL8 to NFS/N1.H7 led to clones (H7 NeoRas.F3, H7 NeoRas.E2) that had incorporated the entire 6.6 Kb human mutant H-RAS genome. The mutant RAS-containing clones demonstrated greater proliferation than parent cells or cells containing a control (neo-resistance) vector over a range of suboptimal IL-3 dose and in optimal IL-3 concentrations had a faster doubling rate than parent cells. The clone H7 NeoRas.F3 was studied biochemically, and found to constitutively form 3-fold more 3H-diacylglycerol than the parent cell line upon exposure to 3H-glycerol. PMA could partially repair the proliferative defect of NFS/N1.H7 compared to the RAS-expressor. These studies affirm a secondary, accelerating role for a mutant RAS gene product acting through pKC to promote clonal expansion of immortal myeloid cells stimulated by IL-3.

Original languageEnglish (US)
Pages (from-to)662-668
Number of pages7
JournalLeukemia
Volume3
Issue number9
StatePublished - Jan 1 1989

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ASJC Scopus subject areas

  • Hematology
  • Oncology
  • Cancer Research

Cite this

Boswell, H. S., Harrington, M. A., Burgess, G. S., Nahreini, T. L., Derigs, H. G., Hodges, T. D., English, D., Crean, C. D., & Gabig, T. G. (1989). A mutant RAS gene acts through protein kinase C to augment interleukin-3 dependent proliferation in a fastidious immortal myeloid cell line. Leukemia, 3(9), 662-668.