Abstract
Receptor activator of NF-κB ligand (RANKL) and interferon gamma (IFN-γ) are critical and opposing mediators of osteoclastogenesis, exerting stimulatory and inhibitory effects, respectively. Cathepsin K (CTSK) is a secreted protease that plays an essential role in osteoclastic bone resorption. We have examined the role of IFN-γ in the regulation of CTSK expression in the murine monocytic RAW 264.7 cell line, which can be readily differentiated to bone-resorbing osteoclasts upon RANKL treatment. Real-time RT-PCR reveals that RANKL stimulates CTSK mRNA expression in a dose- and time-dependent fashion, but that RANKL does not alter the expression of cathepsin L (CTSL) and cathepsin S (CTSS) mRNA. IFN-γ stimulates both CTSL and CTSS expression after 3 days, but fails to significantly alter CTSK expression. IFN-γ markedly inhibits the stimulation of CTSK mRNA and protein by RANKL, whereas RANKL suppresses the stimulation of CTSL and CTSS mRNA by IFN-γ. IFN-γ also ablates the RANKL induced osteoclastic differentiation of RAW cells. In RAW cells stably transfected with a CTSK promoter-luciferase plasmid containing the 1618 bp upstream of the transcription initiation site, IFN-γ inhibits CTSK promoter activity and ablates its induction by RANKL. In conclusion, IFN-γ and RANKL differentially regulate cathepsin K, S, and L gene expression in pre-osteoclastic cells, and there appears to be significant cross talk between the signal transduction pathways mediating the responses to RANKL and IFN-γ.
| Original language | English |
|---|---|
| Pages (from-to) | 756-763 |
| Number of pages | 8 |
| Journal | Biochemical and Biophysical Research Communications |
| Volume | 328 |
| Issue number | 3 |
| DOIs | |
| State | Published - Mar 18 2005 |
| Externally published | Yes |
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Keywords
- Cathepsin K
- Cathepsin L
- Cathepsin S
- Interferon-γ
- Osteoclast
- RANK ligand
ASJC Scopus subject areas
- Biochemistry
- Biophysics
- Molecular Biology
Cite this
RANK ligand and interferon gamma differentially regulate cathepsin gene expression in pre-osteoclastic cells. / Pang, Manhui; Martinez, Ariel F.; Jacobs, Jay; Balkan, Wayne E; Troen, Bruce R.
In: Biochemical and Biophysical Research Communications, Vol. 328, No. 3, 18.03.2005, p. 756-763.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - RANK ligand and interferon gamma differentially regulate cathepsin gene expression in pre-osteoclastic cells
AU - Pang, Manhui
AU - Martinez, Ariel F.
AU - Jacobs, Jay
AU - Balkan, Wayne E
AU - Troen, Bruce R.
PY - 2005/3/18
Y1 - 2005/3/18
N2 - Receptor activator of NF-κB ligand (RANKL) and interferon gamma (IFN-γ) are critical and opposing mediators of osteoclastogenesis, exerting stimulatory and inhibitory effects, respectively. Cathepsin K (CTSK) is a secreted protease that plays an essential role in osteoclastic bone resorption. We have examined the role of IFN-γ in the regulation of CTSK expression in the murine monocytic RAW 264.7 cell line, which can be readily differentiated to bone-resorbing osteoclasts upon RANKL treatment. Real-time RT-PCR reveals that RANKL stimulates CTSK mRNA expression in a dose- and time-dependent fashion, but that RANKL does not alter the expression of cathepsin L (CTSL) and cathepsin S (CTSS) mRNA. IFN-γ stimulates both CTSL and CTSS expression after 3 days, but fails to significantly alter CTSK expression. IFN-γ markedly inhibits the stimulation of CTSK mRNA and protein by RANKL, whereas RANKL suppresses the stimulation of CTSL and CTSS mRNA by IFN-γ. IFN-γ also ablates the RANKL induced osteoclastic differentiation of RAW cells. In RAW cells stably transfected with a CTSK promoter-luciferase plasmid containing the 1618 bp upstream of the transcription initiation site, IFN-γ inhibits CTSK promoter activity and ablates its induction by RANKL. In conclusion, IFN-γ and RANKL differentially regulate cathepsin K, S, and L gene expression in pre-osteoclastic cells, and there appears to be significant cross talk between the signal transduction pathways mediating the responses to RANKL and IFN-γ.
AB - Receptor activator of NF-κB ligand (RANKL) and interferon gamma (IFN-γ) are critical and opposing mediators of osteoclastogenesis, exerting stimulatory and inhibitory effects, respectively. Cathepsin K (CTSK) is a secreted protease that plays an essential role in osteoclastic bone resorption. We have examined the role of IFN-γ in the regulation of CTSK expression in the murine monocytic RAW 264.7 cell line, which can be readily differentiated to bone-resorbing osteoclasts upon RANKL treatment. Real-time RT-PCR reveals that RANKL stimulates CTSK mRNA expression in a dose- and time-dependent fashion, but that RANKL does not alter the expression of cathepsin L (CTSL) and cathepsin S (CTSS) mRNA. IFN-γ stimulates both CTSL and CTSS expression after 3 days, but fails to significantly alter CTSK expression. IFN-γ markedly inhibits the stimulation of CTSK mRNA and protein by RANKL, whereas RANKL suppresses the stimulation of CTSL and CTSS mRNA by IFN-γ. IFN-γ also ablates the RANKL induced osteoclastic differentiation of RAW cells. In RAW cells stably transfected with a CTSK promoter-luciferase plasmid containing the 1618 bp upstream of the transcription initiation site, IFN-γ inhibits CTSK promoter activity and ablates its induction by RANKL. In conclusion, IFN-γ and RANKL differentially regulate cathepsin K, S, and L gene expression in pre-osteoclastic cells, and there appears to be significant cross talk between the signal transduction pathways mediating the responses to RANKL and IFN-γ.
KW - Cathepsin K
KW - Cathepsin L
KW - Cathepsin S
KW - Interferon-γ
KW - Osteoclast
KW - RANK ligand
UR - http://www.scopus.com/inward/record.url?scp=13444280103&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=13444280103&partnerID=8YFLogxK
U2 - 10.1016/j.bbrc.2004.12.005
DO - 10.1016/j.bbrc.2004.12.005
M3 - Article
C2 - 15694411
AN - SCOPUS:13444280103
VL - 328
SP - 756
EP - 763
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
SN - 0006-291X
IS - 3
ER -