Myeloperoxidase and oncogene expression in GM-CSF induced bone marrow differentiation

B. D. Jaffe, D. E. Sabath, G. D. Johnson, L. C. Moscinski, K. R. Johnson, G. Rovera, W. M. Nauseef, Michael B. Prystowsky

Research output: Contribution to journalArticle

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Abstract

DNA synthesis, morphology, specific RNA accumulation and rates of specific protein synthesis in GM-CSF stimulated bone marrow progenitor cells were studied. DNA synthesis increased markedly for 64 hours and then gradually decreased to 5% maximal activity by 160 hours. Morphologic examination 40 to 64 hours after stimulation revealed an increasing proportion of immature myeloid cells. After this proliferative peak, cells differentiated into segmented neutrophils and monocytes/macrophages; only mature forms were present by 160 hours. Accumulation of mRNA for c-myb and c-myc was maximal at 40 hours just prior to maximal [3H]thymidine incorporation, while maximal accumulatio of histone type 3 (H3) was coincident with maximal [3H]thymidine incorporation at 64 hours. As proliferation decreased and differentiation proceeded, levels of mRNA for c-myb and H3 decreased markedly, while levels of RNA for c-myc decreased gradually and remained elevated above day 0 levels. Levels of c-fos mRNA fluctuated slightly during the first 64 hours of culture and increased 13-fold by 160 hours when mature cells were present. Similarly, beta-2 microglobulin mRNA increased steadily to maximal levels at 112 to 160 hours which were 15-fold higher than day 0 levels. Myeloperoxidase (MPO) mRNA was present in maximal amounts at 40 to 64 hours after stimulation with GM-CSF as the number of immature myeloid cells peaked. Immunoprecipitation of MPO from pulse-labeled cell lysates demonstrated a 7-fold rise in synthetic rate of MPO of 64 hours and a 28-fold decline by 160 hours when only 5% immature myeloid cells were present. Thus, MPO protein synthesis closely follows MPO mRNA accumulation. Immuno precipitation of lactoferrin, a marker of myeloid secondary granules, demonstrated a gradual 5-fold increase in synthetic rate as the cells matured. Taken together, these data show that maximal expression of the early myeloid differentiation enzyme myeloperoxidase in GM-CSF stimulated normal bone marrow cells occurs during peak proliferation of immature myeloid cells.

Original languageEnglish (US)
Pages (from-to)167-174
Number of pages8
JournalOncogene
Volume2
Issue number2
StatePublished - 1988
Externally publishedYes

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Granulocyte-Macrophage Colony-Stimulating Factor
Oncogenes
Peroxidase
Bone Marrow
Myeloid Cells
Messenger RNA
Bone Marrow Cells
Histones
Thymidine
RNA
beta 2-Microglobulin
Lactoferrin
DNA
Immunoprecipitation
Monocytes
Proteins
Neutrophils
Stem Cells
Macrophages
Enzymes

ASJC Scopus subject areas

  • Cancer Research
  • Genetics
  • Molecular Biology

Cite this

Jaffe, B. D., Sabath, D. E., Johnson, G. D., Moscinski, L. C., Johnson, K. R., Rovera, G., ... Prystowsky, M. B. (1988). Myeloperoxidase and oncogene expression in GM-CSF induced bone marrow differentiation. Oncogene, 2(2), 167-174.

Myeloperoxidase and oncogene expression in GM-CSF induced bone marrow differentiation. / Jaffe, B. D.; Sabath, D. E.; Johnson, G. D.; Moscinski, L. C.; Johnson, K. R.; Rovera, G.; Nauseef, W. M.; Prystowsky, Michael B.

In: Oncogene, Vol. 2, No. 2, 1988, p. 167-174.

Research output: Contribution to journalArticle

Jaffe, BD, Sabath, DE, Johnson, GD, Moscinski, LC, Johnson, KR, Rovera, G, Nauseef, WM & Prystowsky, MB 1988, 'Myeloperoxidase and oncogene expression in GM-CSF induced bone marrow differentiation', Oncogene, vol. 2, no. 2, pp. 167-174.
Jaffe BD, Sabath DE, Johnson GD, Moscinski LC, Johnson KR, Rovera G et al. Myeloperoxidase and oncogene expression in GM-CSF induced bone marrow differentiation. Oncogene. 1988;2(2):167-174.
Jaffe, B. D. ; Sabath, D. E. ; Johnson, G. D. ; Moscinski, L. C. ; Johnson, K. R. ; Rovera, G. ; Nauseef, W. M. ; Prystowsky, Michael B. / Myeloperoxidase and oncogene expression in GM-CSF induced bone marrow differentiation. In: Oncogene. 1988 ; Vol. 2, No. 2. pp. 167-174.
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abstract = "DNA synthesis, morphology, specific RNA accumulation and rates of specific protein synthesis in GM-CSF stimulated bone marrow progenitor cells were studied. DNA synthesis increased markedly for 64 hours and then gradually decreased to 5{\%} maximal activity by 160 hours. Morphologic examination 40 to 64 hours after stimulation revealed an increasing proportion of immature myeloid cells. After this proliferative peak, cells differentiated into segmented neutrophils and monocytes/macrophages; only mature forms were present by 160 hours. Accumulation of mRNA for c-myb and c-myc was maximal at 40 hours just prior to maximal [3H]thymidine incorporation, while maximal accumulatio of histone type 3 (H3) was coincident with maximal [3H]thymidine incorporation at 64 hours. As proliferation decreased and differentiation proceeded, levels of mRNA for c-myb and H3 decreased markedly, while levels of RNA for c-myc decreased gradually and remained elevated above day 0 levels. Levels of c-fos mRNA fluctuated slightly during the first 64 hours of culture and increased 13-fold by 160 hours when mature cells were present. Similarly, beta-2 microglobulin mRNA increased steadily to maximal levels at 112 to 160 hours which were 15-fold higher than day 0 levels. Myeloperoxidase (MPO) mRNA was present in maximal amounts at 40 to 64 hours after stimulation with GM-CSF as the number of immature myeloid cells peaked. Immunoprecipitation of MPO from pulse-labeled cell lysates demonstrated a 7-fold rise in synthetic rate of MPO of 64 hours and a 28-fold decline by 160 hours when only 5{\%} immature myeloid cells were present. Thus, MPO protein synthesis closely follows MPO mRNA accumulation. Immuno precipitation of lactoferrin, a marker of myeloid secondary granules, demonstrated a gradual 5-fold increase in synthetic rate as the cells matured. Taken together, these data show that maximal expression of the early myeloid differentiation enzyme myeloperoxidase in GM-CSF stimulated normal bone marrow cells occurs during peak proliferation of immature myeloid cells.",
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