The regulation of macrophage colony-stimulating factor (M-CSF) gene expression by phorbol esters and calcium ionophore (A23187) was studied in HL-60 cells. In untreated HL-60 cells, M-CSF transcripts were undetectable, but transcripts were present within 2 h of A23187 treatment or within 8 h of phorbol 12,13-dibutyrate (PdBu) treatment. Concurrent treatment of HL-60 cells with A23187 and cycloheximide (CHX) for 6 h led to a superinduction of message over A23187 treatment alone. But concurrent treatment with PdBu and CHX for 24 h abolished expression of the message normally present after 24 h of PdBu treatment. The role of new protein synthesis in transcriptional regulation of M-CSF was studied by run-on transcription assay. Untreated HL-60 cells or cells treated with CHX alone do not transcribe M-CSF mRNA. However, cells treated with A23187 for 6 h and CHX for 1, 3, or 6 h transcribed more M-CSF than cells treated with A23187 alone for 6 h. CHX also regulates M-CSF expression by message stabilization. The t( 1/2 ) of M-CSF mRNA was 45 min in cells treated with A23187 for 6 h or in cells treated with PdBu for 24 h, was over 2 h in HL-60 cells treated with A23187 for 6 h and CHX for 1 h, and was 3 h in cells treated with PdBu for 24 h and CHX for 1 h. We conclude that M-CSF gene expression can be differentially regulated in HL-60 cells and that new protein synthesis plays an important role in both the transcriptional and posttranscriptional regulation.
|Original language||English (US)|
|Number of pages||5|
|Journal||Journal of Immunology|
|State||Published - 1991|
ASJC Scopus subject areas
- Immunology and Allergy