Effects of gamma‐irradiation on the M‐CSF‐promoter linked to a chloramphenicol aminoacyl transferase reporter gene expressed in a clonal murine bone marrow stromal cell line

Mary Ann Sakakeeny, Jean Leif, Wilma Merrill, Denise Pratt, Elizabeth Romanik, Michael Mckenna, T. J. Fitzgerald, Maureen Harrington, Joel S. Greenberger

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

The effects of cytokines produced by bone marrow stromal cells on closely associated hematopoietic cells constitute a major component of the physiology of the hematopoietic microenvironment. A major cytokine produced by marrow stromal cells is macrophage colony‐stimulating factor (M‐CSF). To determine the effect of gamma‐irradiation on the M‐CSF promoter in bone marrow stromal cells, we selected a clonal cell line from the C3H/HeJ mouse marrow stromal cell line D2XRII and stably transfected a reporter construct containing the murine M‐CSF‐promoter linked to a chloramphenicol aminoacyl transferase (CAT) gene. CAT activity was measured at serial time points after gamma‐irradiation in vitro to doses between 500 and 10,000 cGy at a dose rate of 116 cGy/min. D2XRII marrow stromal cells treated with phorbol myristate acetate (40 m̈g/ml, four h), demonstrated a significant two‐fold increase in CAT activity. In contrast, CAT activity measured immediately, 24 h, 72 h or 1 week after gamma‐irradiation, showed no significant increase or decrease in CAT activity. An increase in CAT activity was detected 48 h after irradiation with cells that received 5,000 cGy. Thus, single fraction gamma‐irradiation of plateau phase bone marrow stromal cells did not decrease M‐CSF‐promoter activity. These results are consistent with prior experimental data demonstrating stable levels of release of M‐CSF protein following gamma‐irradiation of bone marrow stromal cells and imply that the stability of transcription of the gene for this important cytokine is protected from irradiation.

Original languageEnglish (US)
Pages (from-to)87-94
Number of pages8
JournalSTEM CELLS
Volume12
Issue number1
DOIs
StatePublished - 1994

    Fingerprint

Keywords

  • Cytokines
  • Ionizing irradiation
  • M‐CSF

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

Cite this